9-Aminomethyl Minocycline Compounds And Use Thereof In Treating Community-Acquired Bacterial Pneumonia (CABP)

ABSTRACT

The invention disclosed herein provides a method for treating Community-Acquired Bacterial Pneumonia (CABP) using 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereof, in either oral or IV doses or a combination of both.

REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/507,410, filed on Jul. 10, 2019, which is a continuation of U.S.patent application Ser. No. 15/798,573, filed on Oct. 31, 2017, now U.S.Pat. No. 10,383,884, issued Aug. 20, 2019, which claims the benefit ofthe filing dates under 35 U.S.C. § 119(e) of U.S. ProvisionalApplication No. 62/416,010, filed on Nov. 1, 2016; 62/422,843, filed onNov. 16, 2016; 62/480,516, filed on Apr. 2, 2017; 62/500,611, filed onMay 3, 2017; and 62/514,415, filed on Jun. 2, 2017, the entire contentsof each of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

The development of the tetracycline antibiotics was the direct result ofa systematic screening of soil specimens collected from many parts ofthe world for evidence of microorganisms capable of producingbactericidal and/or bacteriostatic compositions. The first of thesenovel compounds was introduced in 1948 under the name chlortetracycline.Two years later, oxytetracycline became available. The elucidation ofthe chemical structure of these compounds confirmed their similarity andfurnished the analytical basis for the production of a third member ofthis group in 1952, tetracycline. A new family of minocycline compounds,without the ring-attached methyl group present in earlier tetracyclines,was prepared in 1957 and became publicly available in 1967; andminocycline was in use by 1972.

Recently, research efforts have focused on developing new tetracyclineantibiotic compositions effective under varying therapeutic conditionsand routes of administration. New tetracycline analogues have also beeninvestigated which may prove to be equal to or more effective than theoriginally introduced minocycline compounds. Examples include U.S. Pat.Nos. 2,980,584; 2,990,331; 3,062,717; 3,165,531; 3,454,697; 3,557,280;3,674,859; 3,957,980; 4,018,889; 4,024,272; and 4,126,680. These patentsare representative of the range of pharmaceutically active tetracyclineand tetracycline analogue compositions.

Historically, soon after their initial development and introduction, thetetracyclines were found to be highly effective pharmacologicallyagainst rickettsia; a number of gram-positive and gram-negativebacteria; and the agents responsible for lymphogranuloma venereum,inclusion conjunctivitis, and psittacosis. Hence, tetracyclines becameknown as “broad spectrum” antibiotics. With the subsequent establishmentof their in vitro antimicrobial activity, effectiveness in experimentalinfections, and pharmacological properties, the tetracyclines as a classrapidly became widely used for therapeutic purposes. However, thiswidespread use of tetracyclines for both major and minor illnesses anddiseases led directly to the emergence of resistance to theseantibiotics even among highly susceptible bacterial species bothcommensal and pathogenic (e.g., Pneumococci and Salmonella). The rise oftetracycline-resistant organisms has resulted in a general decline inuse of tetracyclines and tetracycline analogue compositions asantibiotics of choice. In addition, other antibacterial agents have alsobeen over used creating strains of multiple drug resistant bacteria.Therefore, there is a need for effective antibacterial agents for thetreatment of bacterial infections in general, particularly antibacterialagents with no or less severe resistance by disease-responsiblepathogens.

Community Acquired Bacterial Pneumonia (CABP), also known as CommunityAcquired Pneumonia (CAP) (which terms can be used interchangeably), isdefined as an acute bacterial infection of the pulmonary parenchymaassociated with chest pain, cough, sputum production, difficultybreathing, chills, rigors, fever, or hypotension, and is accompanied bythe presence of a new lobar or multilobar infiltrate on a chestradiograph. Common typical bacterial pathogens that cause CABP includeStreptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus,and Moraxella catarrhalis. Atypical bacterial pathogens such asChlamydophila pneumoniae, Mycoplasma pneumoniae, and Legionellapneumophila also cause CABP.

CABP is a leading cause of morbidity and mortality in the United States(US) and throughout the world (Mandell et al., Clin. Infect. Dis.44:S27-S72, 2007). Four to 6 million cases of CABP occur per year in theUS, resulting in 10 million physician visits, 600,000 hospitalizations,and tens of thousands of deaths. The total cost of CABP to the annual UShealth care budget exceeds $10 billion (in 2007-adjusted dollars)(Niederman et al., Clin. Ther. 20(4): 820-37, 1998). Furthermore, thereis increasing resistance to antibiotics among common pathogens, with aresulting critical need for new antibiotics (Spellberg et al., Clin.Infect. Dis. 46(2):155-164, 2008). Bacterial resistance to the mostfrequently prescribed, currently available antibiotics has limited theirpotential to treat infections, which prevents their use as a first-lineempiric monotherapy. Methicillin-resistant Staphylococcus aureus (MRSA)and multi-drug resistant Streptococcus pneumoniae (MDR-SP) in thecommunity have posed treatment challenges because of resistance topenicillins (resistance rate 100% for both), cephalosporins (100% and11%, respectively, for ceftriaxone), macrolides (83% and 86%,respectively, for azithromycin/erythromycin), and quinolones (73% and2%, respectively, for levofloxacin), in CABP. In addition, the growingconcern about, “collateral damage” associated with use of quinolone andbeta-lactam class antibiotics further underscores the need for newantibiotic treatment options for CABP (Paterson, Clin Infect Dis. 38Suppl 4: S341-345, 2004). Failure of therapy due to resistance willcontinue to contribute to the morbidity and mortality of CABP andtreatment failures of mild disease will result in increasedhospitalizations and contribute to increased healthcare costs.

SUMMARY OF THE INVENTION

The invention described herein provides 9-aminomethyl minocyclines, suchas 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (Omadacycline, or“Compound 1”), for use in the treatment of CABP.

The invention is also partly based on the discovery that 9-aminomethylminocyclines, such as Compound 1, can be provided as IV dosage form forthe treatment of CABP, either alone or in combination with oral dosageform (such as an oral step down after initial IV doses). In certainembodiments, the invention provides the use of 9-aminomethylminocyclines, such as Compound 1, as IV dosage form for the treatment ofCABP.

The invention is further based on the discovery that 9-aminomethylminocyclines, such as Compound 1, has a relatively broad spectrumagainst a wide variety of pathogens associated with CABP, includingintracellular CABP pathogens. The finding that Compound 1 has in vitroactivity against common typical and atypical pathogens, and thesustained epithelial lining fluid (ELF) and alveolar cell (AC) includingalveolar macrophages (AM) concentrations for 24 hours suggest thatCompound 1 has utility as antibacterial agent for the treatment of lowerrespiratory tract bacterial infections caused by susceptible pathogens,such as intracellular CABP pathogens including Legionella pneumophila,Mycoplasma pneumoniae, Chlamydophila pneumoniae, Chlamydophila psittaci,and/or Coxiella burnetii.

The invention is further based on the discovery that 9-aminomethylminocyclines, such as Compound 1, has a relatively broad spectrumagainst a wide variety of pathogens associated with CABP as furtherdescribed below.

9-aminomethyl minocyclines, such as Compound 1, are also advantageousfor treatment of CABP associated antibiotic-resistant pathogens, such asMRSA and penicillin-resistant Streptococcus pneumoniae (PRSP). Due toits efficacy against resistant pathogens, 9-aminomethyl minocyclines,such as Compound 1, can also be used as a front-line therapeutic agentin cases in which known or suspected drug-resistant bacteria may be thecausative pathogen. On the other hand, 9-aminomethyl minocyclines, suchas Compound 1, can also be used as a therapeutic agent in patients whohave previously been treated by other antibiotics, but have hadinadequate response or have developed/exhibited unacceptable orundesirable adverse events (AEs), such as gastrointestinal tracts AEs(GI tract AEs) and/or C. difficile infection.

Thus one aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three intravenous dosesof about 100 mg each, administered 12 hours apart, followed by, (2) oneor more intravenous doses of about 100 mg each, each administered 24hours following the immediate preceding intravenous dose, followed by,(3) one oral dose of about 300 mg, administered in the morning and 12-24hrs after the immediate preceding intravenous dose, followed by, (4) oneor more oral doses of about 300 mg each, each administered 24 hoursfollowing the immediate preceding oral dose, such that the subject istreated.

A related aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three intravenous dosesof about 100 mg each, administered 12 hours apart, followed by, (2)optionally, one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, followed by, (3) optionally, one oral dose of about 300 mg,administered in the morning and 12-24 hrs after the immediate precedingintravenous dose, followed by, (4) optionally, one or more oral doses ofabout 300 mg each, each administered 24 hours following the immediatepreceding oral dose, such that the subject is treated.

A related aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three intravenous dosesof about 100 mg each, administered 12 hours apart, followed by, (2) oneor more intravenous doses of about 100 mg each, each administered 24hours following the immediate preceding intravenous dose, followed by,(3) one or more oral doses of about 300 mg each, each administered 24hours following the immediate preceding dose, such that said subject istreated.

Another related aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three intravenous dosesof about 100 mg each, administered 12 hours apart, followed by, (2)optionally, one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, followed by, (3) one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding dose, such thatthe subject is treated.

In certain embodiments, step (2) consists of one intravenous dose ofabout 100 mg of the subject 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline or salt thereof.

Yes another related aspect of the invention provides a method oftreating Community-Acquired Bacterial Pneumonia (CABP) in a subject inneed of treatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three intravenous dosesof about 100 mg each, administered 12 hours apart, followed by, (2) oneor more intravenous doses of about 100 mg each, each administered 24hours following the immediate preceding intravenous dose, such that thesubject is treated.

In certain embodiments, the method steps are completed within 5 days, 6days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, or 21 days.

In certain embodiments, the steps are completed within 7-14 days, suchas 7-10 days, 11-14 days, or 10 days.

In certain embodiments, the number of days of IV dosing is 3-10 days,such as 3-6 days, 7-10 days, or 5 days.

In certain embodiments, the method comprises one or more oral doses, andwherein the number of days of IV dosing is 4-7 days, such as 4-5 days,6-7 days, or 5 days.

In certain embodiments, the number of days of oral dosing is 1-7 days,such as 1-4 days, 5-7 days, or 5 days.

In certain embodiments, the number of days of IV dosing is 5 days, andthe number of days of oral dosing is 5 days.

Another related aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three oral doses of about300-450 mg each, administered 12 hours apart, followed by, (2)optionally, one or more oral doses of about 300-600 mg each, eachadministered 24 hours following the immediate preceding oral dose, suchthat the subject is treated.

In certain embodiments, each oral dose is about 300 mg.

In certain embodiments, each oral dose is about 450 mg.

In certain embodiments, each oral dose in step (1) is about 300 mg.

In certain embodiments, each oral dose in step (1) is about 450 mg.

In certain embodiments, each oral dose in step (2) is about 300 mg.

In certain embodiments, each oral dose in step (2) is about 450 mg.

In certain embodiments, each oral dose in step (2) is about 600 mg.

In certain embodiments, the first two oral doses of step (1) are each300 mg, and the last oral dose of step (1) is about 300, 450, or 600 mg.

In certain embodiments, the first two oral doses of step (1) are each450 mg, and the last oral dose of step (1) is about 300, 450, or 600 mg.

Another related aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereof,such that the subject is treated, wherein the 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline is administered as once-daily oral dose of300 mg, for 5, 6, 7, or 8 consecutive days.

Another related aspect of the invention provides a method of treatingCommunity-Acquired Bacterial Pneumonia (CABP) in a subject in need oftreatment thereof, comprising administering to the subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) one or two once-dailyoral dose(s) of about 450-600 mg (administered 24 hrs apart for twoonce-daily oral doses), followed by, (2) one or more oral doses of about300-600 mg each, each administered 24 hours following the immediatepreceding oral dose, such that the subject is treated.

In certain embodiments, the dosing regimen is: (1) one or two once-dailyoral dose(s) of about 450 or 600 mg (administered 24 hrs apart for twoonce-daily oral doses), followed by, (2) one or more oral doses of about300 mg each, each administered 24 hours following the immediatepreceding oral dose.

In certain embodiments, the dosing regimen is: (1) two once-daily oraldoses of about 450 mg, administered 24 hrs apart, followed by, (2) oneor more oral doses of about 300 mg each, each administered 24 hoursfollowing the immediate preceding oral dose.

In certain embodiments, the steps are completed within 7 days, 8 days, 9days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17days, 18 days, 19 days, 20 days, or 21 days.

In certain embodiments, the steps are completed within 7-14 days, within7-10 days, within 11-14 days, or within 10 days.

In certain embodiments, the CABP is caused by Staphylococcus aureusincluding methicillin-resistant Staphylococcus aureus (MRSA),Streptococcus pneumoniae including penicillin-resistant Streptococcuspneumoniae (PRSP), Haemophilus influenzae, Moraxella catarrhalis,Klebsiella pneumoniae, Legionella pneumophila, Chlamydophila pneumoniae,Mycoplasma pneumoniae, Chlamydophila psittaci, Coxiella burnetii,Escherichia coli, or a combination thereof.

In certain embodiments, the Streptococcus pneumoniae ispenicillin-resistant Streptococcus pneumoniae (PRSP),macrolide-resistant Streptococcus pneumoniae, cephalosporin-resistantStreptococcus pneumoniae, or multidrug-resistant Streptococcuspneumoniae (MDRSP).

In certain embodiments, the CABP is caused by intracellular pathogens,such as Legionella pneumophila, Mycoplasma pneumoniae, Chlamydophilapneumoniae, Chlamydophila psittaci, Coxiella burnetii, or a combinationthereof.

In certain embodiments, the CABP is caused by Haemophilusparainfluenzae.

In certain embodiments, the subject is a human.

In certain embodiments, each of said oral dose is administeredindependently as two 150-mg tablets.

In certain embodiments, each of said intravenous dose is administeredcontinuously over about 30 minutes (e.g., at least 30 minutes and notmore than 45 minutes).

In certain embodiments, the dosing regimen has a clinical success ratethat is (1) greater than that of moxifloxacin, or (2) within 10% (or12.5%) margin of non-inferiority compared to moxifloxacin, wherein themoxifloxacin is administered as 400 mg intravenous dose once every 24hours for three or more days, followed by one or more doses of 400 mgoral doses of moxifloxacin once every 24 hours.

In certain embodiments, the subject experiences improvement, at day 3 today 5 after step (1), in at least two symptoms selected from: chestpain, frequency or severity of cough, amount of productive sputum, anddifficulty breathing, wherein said symptoms are evaluated on afour-point scale of absent, mild, moderate, and severe, and whereinimprovement is at least a one-point improvement from baseline to theassessment at said day 3 to day 5 (e.g., from severe to moderate, frommoderate to absent, or from mild to absent).

In certain embodiments, the subject, at day 3 to day 5 after step (1),experiences improvement in at least two symptoms and no worsening in anyof the symptoms selected from: chest pain, frequency or severity ofcough, amount of productive sputum, and difficulty breathing, andimprovement in at least one vital sign selected from: body temperature,blood pressure, heart rate, and respiratory rate.

In certain embodiments, the subject undergoes fasting overnight, with nofood or drink except water for at least 6 hours, just before step (3)dosing, and wherein the subject continues fasting after step (3) dosing,with no food for 2 hours, and no dairy products for 4 hours.

In certain embodiments, the salt is a tosylate salt.

In certain embodiments, the method has a clinical success rate of about70%-100%.

In certain embodiments, the clinical success rate is about 75-95%, about80-95%, about 75-90%, about 80-90%, about 75-85%, about 80-85%, about85-90%, about 90-95%, about 80-82%, or about 81%.

In certain embodiments, the clinical success rate is about 75-85%,observed at about 72-120 hours after the administration of the firstintravenous dose.

In certain embodiments, the clinical success rate is about 80-82%, or80% or 81%.

In certain embodiments, the clinical success rate is observed at about5-10 days after the last dose of treatment (e.g., equivalent to a timefor post treatment evaluation in clinically evaluable population, orCE-PTE; or in ITT population).

In certain embodiments, the clinical success rate is about 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, or 97%.

In certain embodiments, the subject has CABP categorized as PORT RiskClass II.

In certain embodiments, the method has a clinical success rate of about70-100%, about 75-96%, about 75-90%, about 80-83%, about 82%, about80-96%, about 90-96%, or about 95%.

In certain embodiments, the clinical success rate is about 75-85%, orabout 90-100%, observed at about 5-10 days after the last dose oftreatment.

In certain embodiments, the clinical success rate is about 82%, or about95%.

In certain embodiments, the subject has CABP categorized as PORT RiskClass III.

In certain embodiments, the method has a clinical success rate of about80-100%, about 85-95%, about 90-95%, about 90-91%, or about 93-94%.

In certain embodiments, the clinical success rate is about 85-100%,observed at about 5-10 days after the last dose of treatment.

In certain embodiments, the clinical success rate is about 90-91%, orabout 93-94%.

In certain embodiments, subject has CABP categorized as PORT Risk ClassIV.

In certain embodiments, the method has a clinical success rate of about70-100%, about 75-95%, about 80-95%, about 83-85%, or about 90-91%.

In certain embodiments, the clinical success rate is about 80-95%,observed at about 5-10 days after the last dose of treatment.

In certain embodiments, the clinical success rate is about 83-85%, orabout 90-91%.

In certain embodiments, the subject has CABP categorized as PORT RiskClass III or IV.

In certain embodiments, the method has a clinical success rate of about75-100%, about 85-95%, about 85-90%, about 88-89%, about 90-95%, orabout 92-93%.

In certain embodiments, the clinical success rate is about 85-95%,observed at about 5-10 days after the last dose of treatment.

In certain embodiments, the clinical success rate is about 88-89%, orabout 92-93%.

In certain embodiments, gastrointestinal (GI) adverse events (AEs)associated with treatment of said subject are mild.

In certain embodiments, GI adverse events (AEs) associated withtreatment of said subject do not result in discontinuation of therapywith the method.

In certain embodiments, treatment of the subject (1) does not result inincreased risk of C. difficile (also known as C. difficile colitis andPseudomembranous colitis) infection in the subject, or (2) does notsubstantially disrupting gut microbiome in the subject.

In certain embodiments, the subject is at risk of, or is predisposed to,developing a C. difficile infection.

In certain embodiments, the subject has recently been treated with oneor more antibiotics (such as broad spectrum antibiotics), has hadsurgery of the gastrointestinal tract, has a disease of the colon (suchas an inflammatory bowel disease or colorectal cancer), has a kidneydisease, has a weakened immune system; is on chemotherapy, haspreviously had C. difficile infection, is 65 years or older, takesproton-pump inhibitors, or is living in an environment that predisposessaid subject to developing C. difficile infection (such as in ahospital, a nursing home, or an assisted living facility).

It should be understood that any one embodiment can be combined with anyother embodiment unless explicitly disclaimed or improper.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 shows the result of mean Compound 1 (Omadacycline) concentrationvs. time profile in AC (Alveolar Cells), plasma, and ELF.

FIG. 2 shows that Compound 1 (“Omadacycline”) demonstrated statisticalnon-inferiority (10% margin) relative to moxifloxacin, for earlyclinical response (ECR) in the ITT (Intent To Treat) population (see thepair of bars on the left) (FDA Primary Endpoint); and for clinicalsuccess at the PTE (Post Treatment/Therapy Evaluation), in both the ITTpopulation (see the middle pair of bars) and the CE-PTE (ClinicallyEvaluable population at the PTE) population (see the right pair of bars)(FDA Secondary Endpoints).

FIG. 3 shows that Compound 1 (“Omadacycline”) demonstrated statisticalnon-inferiority (10% margin) relative to moxifloxacin, based on clinicalsuccess at the PTE in both the ITT population limited to patients withCABP categorized as PORT Risk Class III/IV (see the left pair of bars)and the CE-PTE population limited to patients with CABP categorized asPORT Risk Class III/IV (see the right pair of bars) (EMA co-PrimaryEndpoints).

FIG. 4 shows clinical success rate at PTE for both Compound 1(Omadacycline) and Moxifloxacin, by baseline pathogen in the Compound 1treatment arm with 10 or more isolates. N1=Number of subjects with thespecific baseline pathogen. n=Number of subjects in the specificcategory. Percentages are based on the number of subjects with thespecific baseline pathogen.

FIG. 5 shows plasma concentration versus time curves of omadacyclineafter oral administration. Mean (±SD) plasma concentrations ofomadacycline versus time are shown by omadacycline dose (300, 450, or600 mg) for the pharmacokinetic population. Oral omadacycline doses wereadministered at time 0 on each of 5 consecutive days of dosing in eachof 3 periods. Blood samples were collected for PK analysis on Day 1(left panel) and Day 5 (right panel). Data was pooled by omadacyclinedose for all subjects regardless of the period in which they received aparticular dose.

DETAILED DESCRIPTION OF THE INVENTION

The invention pertains, at least in part, to the discovery that9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (Compound1/Omadacycline) is effective to treat certain bacterial infections, suchas CABP, based on a specific dosage and administration regimen.

The invention is further based on the discovery that 9-aminomethylminocyclines, such as Compound 1, has a relatively broad spectrumagainst a wide variety of pathogens associated with CABP as furtherdescribed below.

In addition, 9-aminomethyl minocyclines, such as Compound 1, also has arelatively broad spectrum against intracellular CABP pathogens. Thefinding that Compound 1 has in vitro activity against common typical andatypical pathogens, and the sustained epithelial lining fluid (ELF) andalveolar cell (AC) including alveolar macrophages (AM) concentrationsfor 24 hours suggest that Compound 1 has utility as antibacterial agentfor the treatment of lower respiratory tract bacterial infections causedby susceptible pathogens, such as intracellular CABP pathogens includingLegionella pneumophila, Mycoplasma pneumoniae, Chlamydophila pneumoniae,Chlamydophila psittaci, and/or Coxiella burnetii.

Thus in one aspect, the invention provides a method of treating CABP ina subject in need of treatment thereof.

In a 1^(st) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (e.g., as atosylate salt) according to the following dosing regimen: (1) threeintravenous (IV) doses of about 100 mg each, administered 12 hoursapart, followed by, (2) one or more IV doses of about 100 mg each, eachadministered 24 hours following the immediate preceding IV dose,followed by, (3) one oral dose of about 300 mg, administered in themorning and 12-24 hrs after the immediate preceding intravenous dose,followed by, (4) one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding oral dose, suchthat the subject is treated.

In a related, 2^(nd) embodiment, the method comprises administering tothe subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100 mg each, administered 12 hours apart, followed by,(2) optionally, one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, followed by, (3) optionally, one oral dose of about 300 mg,administered in the morning and 12-24 hrs after the immediate precedingintravenous dose, followed by, (4) optionally, one or more oral doses ofabout 300 mg each, each administered 24 hours following the immediatepreceding oral dose, such that the subject is treated.

In a related, 3^(rd) embodiment, the method comprises administering tothe subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100-125 mg each, administered 12 hours apart, followedby, (2) optionally, one or more intravenous doses of about 100-125 mgeach, each administered 24 hours following the immediate precedingintravenous dose, followed by, (3) optionally, one oral dose of about300-475 mg (i.e., 3 times of the IV dose administered in oral dose),administered in the morning and 12-24 hrs after the immediate precedingintravenous dose, followed by, (4) optionally, one or more oral doses ofabout 300-475 mg each (i.e., 3 times of the IV dose administered in oraldose), each administered 24 hours following the immediate preceding oraldose, such that the subject is treated. In certain embodiments, the100-125 mg IV dose is about 100-120 mg, about 100-115 mg, about 100-110mg, and about 100-105 mg; and the 300-475 mg oral dose is 3 times of thecorresponding IV dose (i.e., the 300-475 mg oral dose is about 300-360mg, about 300-345 mg, about 300-330 mg, and about 300-315 mg,respectively).

In a 4^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (e.g., as atosylate salt) according to the following dosing regimen: (1) oneintravenous (IV) dose of about 200 mg, followed by, (2) optionally, oneor more IV doses of about 100 mg each, each administered 24 hoursfollowing the immediate preceding IV dose, followed by, (3) optionally,one oral dose of about 300 mg, administered in the morning and 12-24 hrsafter the immediate preceding intravenous dose, followed by, (4) one ormore oral doses of about 300 mg each, each administered 24 hoursfollowing the immediate preceding oral dose, such that the subject istreated.

In a 5^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100 mg each, administered 12 hours apart, followed by,(2) one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, followed by, (3) one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding dose, such thatthe subject is treated.

In a related 6^(th) embodiment, the method comprises administering tothe subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100 mg each, administered 12 hours apart, followed by,(2) optionally, one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, followed by, (3) one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding dose, such thatthe subject is treated.

In a related 7^(th) embodiment, the method comprises administering tothe subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100-125 mg each, administered 12 hours apart, followedby, (2) optionally, one or more intravenous doses of about 100-125 mgeach, each administered 24 hours following the immediate precedingintravenous dose, followed by, (3) one or more oral doses of about300-475 mg (i.e., 3 times of the IV dose administered in oral dose)each, each administered 24 hours following the immediate preceding dose,such that the subject is treated. In certain embodiments, the 100-125 mgIV dose is about 100-120 mg, about 100-115 mg, about 100-110 mg, andabout 100-105 mg; and the 300-475 mg oral dose is 3 times of thecorresponding IV dose (i.e., the 300-475 mg oral dose is about 300-360mg, about 300-345 mg, about 300-330 mg, and about 300-315 mg,respectively).

In an 8^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) one intravenousdose of about 200 mg, followed by, (2) optionally, one or moreintravenous doses of about 100 mg each, each administered 24 hoursfollowing the immediate preceding intravenous dose, followed by, (3)optionally, one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding dose, such thatthe subject is treated.

In a 9^(th) embodiment, step (2) in the method of any one of the 5^(th)to the 8^(th) embodiments consists of one intravenous dose of about 100mg of the subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline orsalt thereof.

In a 10^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100 mg each, administered 12 hours apart, followed by,(2) one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, such that said subject is treated.

In a related 11^(th) embodiment, the method comprises administering tothe subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100-125 mg each, administered 12 hours apart, followedby, (2) one or more intravenous doses of about 100-125 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, such that said subject is treated. In certain embodiments, the100-125 mg IV dose is about 100-120 mg, about 100-115 mg, about 100-110mg, and about 100-105 mg.

In a related 12^(th) embodiment, the method comprises administering tothe subject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) one intravenousdose of about 200 mg, followed by, (2) one or more intravenous doses ofabout 100 mg each, each administered 24 hours following the immediatepreceding intravenous dose, such that said subject is treated.

In a 13^(th) embodiment, the method steps of any one of the1^(st)-12^(th) embodiments are completed within 5 days, 6 days, 7 days,8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16days, 17 days, 18 days, 19 days, 20 days, or 21 days. In certain relatedembodiments, the method steps are completed within 7 days, 8 days, 9days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17days, 18 days, 19 days, or 20 days. In yet another embodiment, themethod steps are completed within 5 days, 6 days, or 21 days. In certainrelated embodiments, the method steps are completed within 5 or 6 days.In certain related embodiments, the method steps are completed within7-14 days.

In a 14^(th) embodiment, the method steps of any one of the1^(st)-12^(th) embodiments are completed within 7-14 days, such as 7-10days, 11-14 days, or 10 days.

In a 15^(th) embodiment, the number of days of IV dosing in the 14^(th)embodiment is 3-10 days, such as 3-6 days, 7-10 days, or 5 days.

In a 16^(th) embodiment, the method of any one of the 1^(st)-15^(th)embodiments comprise one or more oral doses, and wherein the number ofdays of IV dosing is 4-7 days, such as 4-5 days, 6-7 days, or 5 days.

In a 17^(th) embodiment, the number of days of oral dosing in the16^(th) embodiment is 1-7 days, such as 1-4 days, 5-7 days, or 5 days.

In an 18^(th) embodiment, the number of days of IV dosing in any one ofthe 1^(st)-17^(th) embodiments is 5 days, and the number of days of oraldosing is 5 days.

In a 19^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (e.g., as atosylate salt) according to the following dosing regimen: (1) three oraldoses of about 300-450 mg (such as 300 mg or 450 mg) each, administered12 hours apart, followed by, (2) optionally, one or more oral doses ofabout 300-600 mg (such as 300 mg, 450 mg, or 600 mg) each, eachadministered 24 hours following the immediate preceding oral dose, suchthat the subject is treated.

In a 20^(th) embodiment, each oral dose of the 19^(th) embodiment isabout 300 mg.

In a 21^(st) embodiment, each oral dose of the 19^(th) embodiment isabout 450 mg.

In a 22^(nd) embodiment, each oral dose in step (1) of the 19^(th)embodiment is about 300 mg.

In a 23^(rd) embodiment, each oral dose in step (1) of the 19^(th)embodiment is about 450 mg.

In a 24^(th) embodiment, each oral dose in step (2) of the 19^(th),22^(nd), or 23^(rd) embodiment is about 300 mg.

In a 25^(th) embodiment, each oral dose in step (2) of the 19^(th),22^(nd), or 23^(rd) embodiment is about 450 mg.

In a 26^(th) embodiment, each oral dose in step (2) of the 19^(th),22^(nd), or 23^(rd) embodiment is about 600 mg.

In a 27^(th) embodiment, the first two oral doses of step (1) of the19^(th) embodiment are each 300 mg, and the last oral dose of step (1)is about 300, 450, or 600 mg.

In a 28^(th) embodiment, the first two oral doses of step (1) of the19^(th) embodiment are each 450 mg, and the last oral dose of step (1)is about 300, 450, or 600 mg.

In a 29^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (e.g., as atosylate salt) according to the following dosing regimen: (1) one or twoonce-daily oral dose(s) of about 450-600 mg (such as 450 mg, or 600 mg)(administered 24 hrs apart for two once-daily oral doses), followed by,(2) one or more oral doses of about 300-600 mg (such as 300 mg, 450 mg,or 600 mg) each, each administered 24 hours following the immediatepreceding oral dose, such that the subject is treated.

In a 30^(th) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline (e.g., as atosylate salt) according to the following dosing regimen: (1) one or twoonce-daily oral dose(s) of about 300-600 mg (such as 300 mg, 450 mg, or600 mg) (administered 24 hrs apart for two once-daily oral doses),followed by, (2) one or more oral doses of about 300-600 mg (such as 300mg, 450 mg, or 600 mg) each, each administered 24 hours following theimmediate preceding oral dose, such that the subject is treated.

In a 31^(st) embodiment, the method comprises administering to thesubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof, such that the subject is treated, wherein the9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline is administered asonce-daily oral dose of 300 mg, for 5, 6, 7, or 8 consecutive days.

In a 32^(nd) embodiment, the dosing regimen of the 29^(th) embodimentis: (1) one or two once-daily oral dose(s) of about 450-600 mg, such as450 mg or 600 mg, (administered 24 hrs apart for two once-daily oraldoses), followed by, (2) one or more oral doses of about 300 mg each,each administered 24 hours following the immediate preceding oral dose.

In a 33^(rd) embodiment, the dosing regimen of the 29^(th) embodimentis: (1) two once-daily oral doses of about 450 mg, administered 24 hrsapart, followed by, (2) one or more oral doses of about 300 mg each,each administered 24 hours following the immediate preceding oral dose.

In a 34^(th) embodiment, the dosing regimen of the 29^(th) embodimentis: (1) two once-daily oral doses of about 600 mg, administered 24 hrsapart, followed by, (2) one or more oral doses of about 300 mg each,each administered 24 hours following the immediate preceding oral dose.

In a 35^(th) embodiment, the steps of any one of the 25^(th)-34^(th)embodiments are completed within 7 days, 8 days, 9 days, 10 days, 11days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19days, 20 days, or 21 days.

In a 36^(th) embodiment, the steps of any one of the 25^(th)-34^(th)embodiments are completed within 7-14 days, within 7-10 days, within11-14 days, or within 10 days.

In a 37^(th) embodiment, the CABP of any one of the 1^(st)-36^(th)embodiments is caused by Staphylococcus aureus includingmethicillin-resistant Staphylococcus aureus (MRSA), Streptococcuspneumoniae including penicillin-resistant Streptococcus pneumoniae(PRSP), Haemophilus influenzae, Moraxella catarrhalis, Klebsiellapneumoniae, Legionella pneumophila, Chlamydophila pneumoniae, Mycoplasmapneumoniae, Chlamydophila psittaci, Coxiella burnetii, Escherichia coli,or a combination thereof.

In a 38^(th) embodiment, the Streptococcus pneumoniae of the 37^(th)embodiment is penicillin-resistant Streptococcus pneumoniae (PRSP),macrolide-resistant Streptococcus pneumoniae, cephalosporin-resistantStreptococcus pneumoniae, or multidrug-resistant Streptococcuspneumoniae (MDRSP).

In a 39^(th) embodiment, the CABP of any one of the 1^(st)-36^(th)embodiments is caused by intracellular pathogens, such as Legionellapneumophila, Mycoplasma pneumoniae, Chlamydophila pneumoniae,Chlamydophila psittaci, Coxiella burnetii, or a combination thereof.

In a 40^(th) embodiment, the CABP of any one of the 1^(st)-36^(th)embodiments is caused by Haemophilus parainfluenzae.

In a 41^(st) embodiment, the subject of any of the 1^(st)-40^(th)embodiments is a human.

In a 42^(nd) embodiment, each of the oral dose of any of the1^(st)-41^(st) embodiments is administered independently as two 150-mgtablets.

In a 43^(rd) embodiment, each of the intravenous dose of any of the1^(st)-42^(nd) embodiments is administered continuously over about 30minutes (e.g., at least 30 minutes and not more than 45 minutes).

In a 44^(th) embodiment, the dosing regimen of any of the 1^(st)-43^(rd)embodiments has a clinical success rate that is within 10% (or 12.5%)margin of non-inferiority compared to moxifloxacin administered as 400mg intravenous dose once every 24 hours for three or more days, followedby one or more doses of 400 mg oral doses of moxifloxacin once every 24hours.

In a 45^(th) embodiment, the subject of any of the 1^(st)-44^(th)embodiments: (1) has at least 3 of the symptoms selected from: cough,production of purulent sputum, dyspnea (shortness of breath), andpleuritic chest pain; (2) has at least two abnormal vital signs selectedfrom: fever or hypothermia (temperature >38.0° C. [100.4° F.] or <36.0°C. [95.5° F.]), hypotension with systolic blood pressure (SBP)<90 mm Hg,tachycardia (heart rate >90 beats per minute (bpm)), and tachypnea(respiratory rate (RR)>20 breaths/minute); (3) has at least one clinicalsign or laboratory finding associated with CABP: hypoxemia (partialpressure of arterial oxygen [PaO₂] <60 mm Hg by arterial blood gas [ABG]or oxygen saturation <90% by pulse oximetry), clinical evidence (e.g.,by physical examination findings) of pulmonary consolidation (e.g.,dullness on percussion, bronchial breath sounds, or egophony), and anelevated total white blood cell (WBC) count (>12,000 cells/mm³) orleucopenia (WBC<4,000 cells/mm³) or elevated immature neutrophils (>15%band forms, regardless of total peripheral WBC count); (4) hasradiographically-confirmed pneumonia (i.e., new or progressive pulmonaryinfiltrate(s) in a lobar or a multilobar distribution on chest X-ray(CXR) or chest computed tomography (CT) scan consistent with acutebacterial pneumonia within 24 or 48 hours prior to step (1)); and (5)has disease categorized as being PORT Risk Class II, III, or IV, or hasappropriate sputum specimen characterized by fewer than 10 squamousepithelial cells and more than 25 polymorphonuclear cells per low powerfield.

In a 46^(th) embodiment, the subject of any of the 1^(st)-44^(th)embodiments experience improvement, at day 3 to day 5 after step (1), inat least two symptoms selected from: chest pain, frequency or severityof cough, amount of productive sputum, and difficulty breathing, whereinthe symptoms are evaluated on a four-point scale of absent, mild,moderate, and severe, and wherein improvement is at least a one-pointimprovement from baseline to the assessment at the day 3 to day 5 (e.g.,from severe to moderate, from moderate to absent, or from mild toabsent).

In a 47^(th) embodiment, the subject of any of the 1^(st)-46^(th)embodiments, at day 3 to day 5 after step (1), experience improvement inat least two symptoms and no worsening in any of the symptoms selectedfrom: chest pain, frequency or severity of cough, amount of productivesputum, and difficulty breathing, and improvement in at least one vitalsign selected from: body temperature, blood pressure, heart rate, andrespiratory rate.

Compound 1 was found to have a significant food effect, in that foodconsumption has a significant impact on the oral bioavailability oforally administered 300 mg dose of Compound 1. See Example 3. A PK studyin healthy volunteers showed that, compared with a fasted dose,bioavailability was reduced by 15% to 17% for a nondairy meal 4 hoursbefore dosing, 40% to 42% for a nondairy meal 2 hours before dosing, and59% to 63% for a dairy meal 2 hours before dosing. Thus, the effect offood was more pronounced when a high-fat meal was consumed closer todosing and when dairy was included in the meal. Based on this result,oral Compound 1 should be administered at least 6 hours following a mealin order to achieve maximum bioavailability for the oral dose designedto achieve therapeutic efficacy.

Thus in a 48^(th) embodiment, the subject of any one of thefirst-47^(th) embodiments undergoes fasting overnight, with no food ordrink except water for at least 6 hours, just before step (3) dosing,and wherein the subject continues fasting after step (3) dosing, with nofood for 2 hours, and no dairy products for 4 hours.

In a 49^(th) embodiment, the salt of any one of the 1^(st)-48^(th)embodiments is a tosylate salt.

In a 50^(th) embodiment, the method of any one of the 1^(st)-49^(th)embodiments has a clinical success rate of about 70%-100%.

In a 51^(st) embodiment, the clinical success rate of the 50^(th)embodiment is about 75-95%, about 80-95%, about 75-90%, about 80-90%,about 75-85%, about 80-85%, about 85-90%, about 90-95%, about 80-82%, orabout 81%.

In a 52^(nd) embodiment, the clinical success rate of the 51^(st)embodiment is about 75-85%, observed at about 72-120 hours after theadministration of the first intravenous dose.

In a 53^(rd) embodiment, the clinical success rate of the 52^(nd)embodiment is about 80-82%, or 80% or 81%.

In a 54^(th) embodiment, the clinical success rate of the 51^(st)embodiment is observed at about 5-10 days after the last dose oftreatment (e.g., equivalent to a time for post treatment evaluation inclinically evaluable population, or CE-PTE; or in ITT population).

In a 55^(th) embodiment, the clinical success rate of the 54^(th)embodiment is about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97%.

In a 56^(th) embodiment, the subject of any one of the 1^(st)-55^(th)embodiments has CABP categorized as PORT Risk Class II.

In a 57^(th) embodiment, the method of the 56^(th) embodiment has aclinical success rate of about 70-100%, about 75-96%, about 75-90%,about 80-83%, about 82%, about 80-96%, about 90-96%, or about 95%.

In a 58^(th) embodiment, the clinical success rate of the 57^(th)embodiment is about 75-85%, or about 90-100%, observed at about 5-10days after the last dose of treatment.

In a 59^(th) embodiment, the clinical success rate of the 58^(th)embodiment is about 82%, or about 95%.

In a 60^(th) embodiment, the subject of and one of the 1^(st)-55^(th)embodiments has CABP categorized as PORT Risk Class III.

In a 61^(st) embodiment, the method of any one of the 1^(st)-60^(th)embodiments has a clinical success rate of about 80-100%, about 85-95%,about 90-95%, about 90-91%, or about 93-94%.

In a 62^(nd) embodiment, the clinical success rate of the 61^(st)embodiment is about 85-100%, observed at about 5-10 days after the lastdose of treatment.

In a 63^(rd) embodiment, the clinical success rate of the 62^(nd)embodiment is about 90-91%, or about 93-94%.

In a 64^(th) embodiment, the subject of any one of the 1^(st)-55^(th)embodiments has CABP categorized as PORT Risk Class IV.

In a 65^(th) embodiment, the method of the 64^(th) embodiment has aclinical success rate of about 70-100%, about 75-95%, about 80-95%,about 83-85%, or about 90-91%.

In a 66^(th) embodiment, the clinical success rate of the 65^(th)embodiment is about 80-95%, observed at about 5-10 days after the lastdose of treatment.

In a 67^(th) embodiment, the clinical success rate of the 66^(th)embodiment is about 83-85%, or about 90-91%.

In a 68^(th) embodiment, the subject of any one of the 1^(st)-55^(th)embodiments has CABP categorized as PORT Risk Class III or IV.

In a 69^(th) embodiment, the method of the 68^(th) embodiment has aclinical success rate of about 75-100%, about 85-95%, about 85-90%,about 88-89%, about 90-95%, or about 92-93%.

In a 70^(th) embodiment, the clinical success rate of the 69^(th)embodiment is about 85-95%, observed at about 5-10 days after the lastdose of treatment.

In a 71^(st) embodiment, the clinical success rate of the 70^(th)embodiment is about 88-89%, or about 92-93%.

In a 72^(nd) embodiment, gastrointestinal (GI) adverse events (AEs)associated with treatment of the subject in method of any one of the1^(st)-71^(st) embodiments are mild.

In a 73^(rd) embodiment, GI adverse events (AEs) associated withtreatment of the subject in method of any one of the 1^(st)-71^(st)embodiments do not result in discontinuation of therapy with the method.

In a 74^(th) embodiment, treatment of the subject in the method of anyone of the 1^(st)-73^(rd) embodiments (1) does not result in increasedrisk of C. difficile (also known as C. difficile colitis andPseudomembranous colitis) infection in the subject, or (2) does notsubstantially disrupting gut microbiome in the subject.

In a 75^(th) embodiment, the subject of the 74^(th) embodiment is atrisk of, or is predisposed to, developing a C. difficile infection.

In a 76^(th) embodiment, the subject of the 75^(th) embodiment hasrecently been treated with one or more antibiotics (such as broadspectrum antibiotics), has had surgery of the gastrointestinal tract,has a disease of the colon (such as an inflammatory bowel disease orcolorectal cancer), has a kidney disease, has a weakened immune system;is on chemotherapy, has previously had C. difficile infection, is 65years or older, takes proton-pump inhibitors, or is living in anenvironment that predisposes the subject to developing C. difficileinfection (such as in a hospital, a nursing home, or an assisted livingfacility).

In a 77^(th) embodiment, in any of the preceding methods, GI adverseevents (AEs) associated with treatment of the subject do not result indiscontinuation of therapy with the method, and treatment of the subject(1) does not result in increased risk of C. difficile (e.g., C.difficile colitis and Pseudomembranous colitis) infection in the subjector (2) does not substantially disrupting gut microbiome in the subject.

As used herein, the term “subject” may include animals (e.g., non-humanmammal) capable of suffering from a bacterial infection. Examples ofsubjects include animals such as farm animals (e.g., cows, pigs, horses,goats, rabbits, sheep, etc.), lab animals (mice, rats, etc.), pets(e.g., dogs, cats, ferrets, etc.), and primates (e.g., humans andnon-human primates such as monkeys, gorillas, chimpanzees, etc.).

In any of the above embodiments, the subject may be a human, a non-humanprimate, or a non-human mammal.

The term “treating” or “treatment” refers to the amelioration,eradication, or diminishment of one or more symptoms of the disorder,e.g., a bacterial infection, to be treated. In certain embodiments, thedisorder term includes the eradication of bacteria associated with theinfection to be treated.

The term “prophylaxis” means to prevent or reduce the risk of bacterialinfection.

The term “resistance” or “resistant” refers to the antibiotic/organismstandards as defined by the Clinical and Laboratories StandardsInstitute (CLSI) and/or the Food and Drug Administration (FDA).

In certain embodiments, the infection may be resistant to otherantibiotics, such as penicillin or tetracycline.

The term “effective amount” includes the amount of the tetracyclinecompound (e.g., Compound 1) needed to treat a bacterial infection (e.g.,CABP). For example, an effective amount describes an efficacious levelsufficient to achieve the desired therapeutic effect through the killingof bacteria and/or inhibition of bacterial growth. Preferably, thebacterial infection is treated when the pathogen (e.g., bacteria) iseradicated. The bacterial infection is also treated when at least onesymptom of infection is reduced, alleviated, or eliminated.

The term “evaluable clinical success” refers to a clinical trialparticipant who: (1) did not meet any criteria for evaluable clinicalfailure; (2) did not receive potentially effective non-study antibioticsfor any other reason; and (3) the blinded evaluator indicated at thetest of cure evaluation that the infection had sufficiently resolvedsuch that antibiotics were not needed.

The term “evaluable clinical failure” refers to a clinical trialparticipant who met any one of the following criteria: the blindedevaluator discontinued study drug and indicated that the infection hadresponded inadequately such that alternative antibiotic(s) were needed;the blinded evaluator discontinued study drug because of an adverseevent that was assessed as probably or possibly drug-related; theprimary site of infection was surgically removed; or the subject had noevaluation after the end of intravenous or oral treatment.

The term “clinical success rate” refers to the number of evaluableclinical successes divided by the total number of population in thetrial.

The term “microbiologically evaluable clinical success rate” refers tothose who met the definition of evaluable clinical success and had aninfecting pathogen at baseline.

In one embodiment, the effective amount of the tetracycline compound,e.g. 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline, whenadministered orally is from about 150 to about 600 mg, about 150 toabout 450 mg, from about 150 to about 300 mg, or about 300 mg.

In certain embodiments, each oral dose is administered as multiples of150 mg doses (e.g., 150 mg, 2×150 mg, 3×150 mg, or 4×150 mg). Forexample, a 300 mg oral dose may consists of two 150 mgtablets/pills/capsules/gels, etc.

In another embodiment, the effective amount of the tetracyclinecompound, e.g., 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline, whenadministered intravenously (IV) is from about 50 to about 200 mg, fromabout 50 to about 150 mg, from about 50 to about 100 mg, or about 100mg, or about 200 mg.

The compound, either in IV formulation or in oral formulation, may beadministered as a salt (e.g., tosylate salt or hydrochloride salt) or asa free base. For example, any salt or a polymorph of a salt, such as atosylate salt of Compound 1, as described in U.S. Pat. No. 8,383,610(incorporated herein by reference), may be used in the instantinvention. In addition, any formulation, such as oral formulation intablet form, as described in U.S. Pat. No. 9,314,475 (incorporatedherein by reference), may be used in the instant invention.

It is to be understood that wherever values and ranges are providedherein, e.g., in ages of subject populations, dosages, and blood levels,all values and ranges encompassed by these values and ranges, includingrecited upper and/or lower limits of the ranges, are meant to beencompassed within the scope of the present invention. Moreover, allvalues in these values and ranges may also be the upper or lower limitsof a range.

In another embodiment, the tetracycline compound (e.g., Compound 1) maybe administered once or twice per day, either intravenously or orally.In certain embodiments, twice per day administration has two equaldoses.

In certain embodiments, the 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline has a microbiologically evaluable clinicalsuccess rate of greater than about 60%. In certain embodiments, thecompound of the invention has a clinical success rate of greater thanabout 60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 93.7%, 94%, 95%, 96%, 97%,97.5%, 98%, 99% or more, either in the intent to treat (ITT) patientpopulation or in the clinically evaluable (CE) patient population.

As used herein, an “Intent-to-Treat (or ITT)” population refers to allenrolled clinical trial subjects. In certain embodiments, the ITTpopulation is further limited to all enrolled clinical trial subjectswho have received at least one dose of the study drug (e.g., Compound1). A “Clinically Evaluable (or CE)” population refers to all ITTsubjects who had a qualifying infection as defined by the relevantclinical protocol, e.g., those with CABP. “Clinical success” refers tothe continued improvement or complete resolution of baseline symptoms inthe ITT or CE populations, assessed by the clinical investigator, at aset period (e.g., 10 to 17 days) after the last dose of the study drug.

In one example, a subject is treated intravenously followed by an oralstep down. In certain embodiments, the subject is treated directly byoral dose without any preceding IV dose.

In certain embodiments, the present invention provides a method oftreating a subject for an infection (e.g., CABP), comprisingadministering to the subject an effective amount of Compound 1 or a saltthereof wherein the subject is initially treated about 1, 2, 3, 4, or 5days intravenously, followed by about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, or 15 days of oral treatment, such that the subject istreated. In certain embodiments, the first day of IV treatment consistsof a higher loading dose (e.g., 2× dose, or 2×100 mg doses). In certainembodiments, each IV dose from day 2 and beyond is administered about 24hrs from the immediate preceding IV dose. In certain embodiments, thetotal treatment period is about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,or 15 days. In certain embodiments, one (e.g., the 2^(nd) IV loadingdose) or both of the IV loading doses is replaced by a 300 mg or 450 mgoral dose.

In another embodiment, the present invention provides a method oftreating a subject for an infection (e.g., CABP), comprisingadministering to said subject an effective amount of Compound 1 or asalt thereof wherein the subject initially treated intravenously haselevated compound 1 blood levels followed by reduced Compound 1 bloodlevels with oral treatment, such that the subject is treated. In certainembodiments, the initially elevated compound 1 blood level is achievedby a higher (e.g., 2×) loading dose(s), such as 2 IV loading doses ofabout 100 mg each.

Pharmaceutical Compositions of the Invention

The invention also utilizes or pertains to pharmaceutical compositionscomprising a therapeutically effective amount of a tetracycline compound(e.g., a 9-aminomethyl tetracycline compound, e.g.,9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline) or a salt thereofand, optionally, a pharmaceutically acceptable carrier.

In a further embodiment, the invention pertains to a pharmaceuticalcomposition comprising from about 100 to about 700 mg (e.g., about 300,450, or 600 mg) of 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline ora salt thereof and a pharmaceutically acceptable carrier. In a furtherembodiment, the pharmaceutically acceptable carrier is acceptable fororal administration. In another further embodiment, the tetracyclinecompound is a free base or a tosylate salt.

In yet another further embodiment, the composition comprises from about110 to about 490 mg, from about 120 to about 480 mg, from about 130 toabout 470 mg, from about 140 to about 460 mg, from about 150 to about450 mg, from about 160 to about 440 mg, from about 170 mg to about 430mg, from about 180 mg to about 420 mg, from about 190 mg to about 410mg, from about 200 mg to about 400 mg, from about 210 mg to about 390mg, from about 220 mg to about 380 mg, from about 230 mg to about 370mg, from about 240 mg to about 360 mg, from about 250 mg to about 350mg, from about 260 mg to about 340 mg, from about 270 mg to about 330mg, from about 280 mg to about 320 mg, from about 290 mg to about 310mg, or about 300 mg of 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline. Optionally, the pharmaceutically acceptablecarrier is suitable for oral administration.

In another embodiment, the invention also pertains to a pharmaceuticalcomposition comprising from about 50 to about 250 mg (e.g., about 100mg) of 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline or a saltthereof (e.g., a hydrochloride salt) and a pharmaceutically acceptablecarrier suitable for intravenous administration.

In yet another further embodiment, the composition comprises from about100 to about 300 mg, from about 125 to about 275 mg, from about 150 mgto about 250 mg, from about 100 mg to about 200 mg, about 100 mg, orabout 200 mg of IV or oral 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline.

The language “pharmaceutically acceptable carrier” includes substancescapable of being co-administered with the tetracycline compound of theinvention, e.g., 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline, andwhich allow the tetracycline compound to perform its intended function,e.g., treat or prevent a bacterial infection. Suitable pharmaceuticallyacceptable carriers include but are not limited to water, saltsolutions, alcohol, vegetable oils, polyethylene glycols, gelatin,lactose, amylose, magnesium stearate, talc, silicic acid, viscousparaffin, perfume oil, fatty acid monoglycerides and diglycerides,petroethral fatty acid esters, hydroxymethyl-cellulose,polyvinylpyrrolidone, etc. The pharmaceutical preparations can besterilized and if desired mixed with auxiliary agents, e.g., lubricants,preservatives, stabilizers, wetting agents, emulsifiers, salts forinfluencing osmotic pressure, buffers, colorings, flavorings and/oraromatic substances and the like which do not deleteriously react withthe active compounds of the invention.

The tetracycline compounds of the invention (e.g., Compound 1) that arebasic in nature are capable of forming a wide variety of salts withvarious inorganic and organic acids. The acids that may be used toprepare pharmaceutically acceptable acid addition salts of theminocycline compounds of the invention that are basic in nature arethose that form nontoxic acid addition salts, i.e., salts containingpharmaceutically acceptable anions, such as the hydrochloride,hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acidphosphate, isonicotinate, acetate, lactate, salicylate, citrate, acidcitrate, tartrate, pantothenate, bitartrate, ascorbate, succinate,maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate,formate, benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate and palmoate [i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)] salts. Although such saltsmust be pharmaceutically acceptable for administration to a subject,e.g., a mammal, it is often desirable in practice to initially isolate aminocycline compound of the invention from the reaction mixture as apharmaceutically unacceptable salt and then simply convert the latterback to the free base compound by treatment with an alkaline reagent andsubsequently convert the latter free base to a pharmaceuticallyacceptable acid addition salt. The acid addition salts of the basecompounds of this invention are readily prepared by treating the basecompound with a substantially equivalent amount of the chosen mineral ororganic acid in an aqueous solvent medium or in a suitable organicsolvent, such as methanol or ethanol. Upon careful evaporation of thesolvent, the desired solid salt is readily obtained. Preferably, thetetracycline compound of the invention is administered as a tosylate(e.g., p-toluenesulfonate) salt or as a freebase orally or as ahydrochloride salt intravenously.

The tetracycline compounds of the invention (e.g., Compound 1) andpharmaceutically acceptable salts thereof can be administered via eitherthe oral, parenteral or topical routes. In general, these compounds aremost desirably administered in effective dosages, depending upon theweight and condition of the subject being treated and the particularroute of administration chosen. Variations may occur depending upon thespecies of the subject being treated and its individual response to saidmedicament, as well as on the type of pharmaceutical formulation chosenand the time period and interval at which such administration is carriedout.

The pharmaceutical compositions of the invention may be administeredalone or in combination with other known compositions for treatingtetracycline responsive states in a subject, e.g., a mammal. Mammalsinclude pets (e.g., cats, dogs, ferrets, etc.), farm animals (cows,sheep, pigs, horses, goats, etc.), lab animals (rats, mice, monkeys,etc.), and primates (chimpanzees, humans, gorillas). The language “incombination with” a known composition is intended to includesimultaneous administration of the composition of the invention and theknown composition, administration of the composition of the inventionfirst, followed by the known composition and administration of the knowncomposition first, followed by the composition of the invention. Any ofthe therapeutic compositions known in the art for treating tetracyclineresponsive states can be used in the methods of the invention.

The compounds of the invention may be administered alone or incombination with pharmaceutically acceptable carriers or diluents by anyof the routes previously mentioned, and the administration may becarried out in single or multiple doses. For example, the noveltherapeutic agents of this invention can be administered advantageouslyin a wide variety of different dosage forms, i.e., they may be combinedwith various pharmaceutically acceptable inert carriers in the form oftablets, capsules, lozenges, troches, hard candies, powders, sprays,creams, salves, suppositories, jellies, gels, pastes, lotions,ointments, aqueous suspensions, injectable solutions, elixirs, syrups,and the like. Such carriers include solid diluents or fillers, sterileaqueous media and various non-toxic organic solvents, etc. Moreover,oral pharmaceutical compositions can be suitably sweetened and/orflavored. In general, the therapeutically-effective tetracyclinecompounds of this invention are present in such dosage forms atconcentration levels ranging from about 5.0% to about 70% by weight.

For oral administration, tablets containing various excipients such asmicrocrystalline cellulose, sodium citrate, calcium carbonate, dicalciumphosphate and glycine may be employed along with various disintegrantssuch as starch (and preferably corn, potato or tapioca starch), alginicacid and certain complex silicates, together with granulation binderslike polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,lubricating agents such as magnesium stearate, sodium lauryl sulfate andtalc are often very useful for tableting purposes. Solid compositions ofa similar type may also be employed as fillers in gelatin capsules;preferred materials in this connection also include lactose or milksugar as well as high molecular weight polyethylene glycols.

When aqueous suspensions and/or elixirs are desired for oraladministration, the active ingredient may be combined with varioussweetening or flavoring agents, coloring matter or dyes, and, if sodesired, emulsifying and/or suspending agents as well, together withsuch diluents as water, ethanol, propylene glycol, glycerin and variouslike combinations thereof.

For parenteral administration (including intraperitoneal, subcutaneous,intravenous, intradermal or intramuscular injection), solutions of atherapeutic compound of the present invention in either sesame or peanutoil or in aqueous propylene glycol may be employed. The aqueoussolutions should be suitably buffered (preferably pH greater than 8) ifnecessary and the liquid diluent first rendered isotonic.

These aqueous solutions are suitable for intravenous injection purposes.The oily solutions are suitable for intraarticular, intramuscular andsubcutaneous injection purposes. The preparation of all these solutionsunder sterile conditions is readily accomplished by standardpharmaceutical techniques well known to those skilled in the art. Forparenteral application, examples of suitable preparations includesolutions, preferably oily or aqueous solutions as well as suspensions,emulsions, or implants, including suppositories. Therapeutic compoundsmay be formulated in sterile form in multiple or single dose formatssuch as being dispersed in a fluid carrier such as sterile physiologicalsaline or 5% saline dextrose solutions commonly used with injectables.

For enteral application, particularly suitable are tablets, dragees orcapsules having talc and/or carbohydrate carrier binder or the like, thecarrier preferably being lactose and/or corn starch and/or potatostarch. A syrup, elixir or the like can be used wherein a sweetenedvehicle is employed. Sustained release compositions can be formulatedincluding those wherein the active component is protected withdifferentially degradable coatings, e.g., by microencapsulation,multiple coatings, etc.

In addition to treatment of human subjects, the therapeutic methods ofthe invention also will have significant veterinary applications, e.g.for treatment of livestock such as cattle, sheep, goats, cows, swine andthe like; poultry such as chickens, ducks, geese, turkeys and the like;horses; and pets such as dogs and cats. Also, the compounds of theinvention may be used to treat non-animal subjects, such as plants.

EXEMPLIFICATION OF THE INVENTION Example 1 An Open-Label, ParallelGroup, Multiple IV Dose Study to Assess Intra-Pulmonary Steady-StateConcentrations of Compound 1 and Tigecycline in Healthy Adult Subjects

To be effective in lower respiratory tract infections (RTI), anantibiotic must attain adequate concentrations in respiratory tissue toaffect respiratory pathogens. Both extracellular and intracellularpathogens may cause infection, and therefore extracellular andintracellular concentrations of antibiotics must be adequate to coverall of the pathogens. The concentration of antibiotic in bronchialmucosa provides a reliable guide to bronchial penetration of the drugand may be a better predictor of clinical efficacy than serum levels fortreatment of bronchitis and bronchopneumonia.

Epithelial lining fluid (ELF) and alveolar cells (AC), including mostlyalveolar macrophages (AM), have been advocated as important infectionsites for common extracellular and intracellular pathogens,respectively. Direct measurement of the concentration of antimicrobialagents in the ELF allows for a more informed approach to appropriatedosing of the agent for RTI, and to evaluate the pharmacokinetic (PK)and exposure-response targets of the drug for respiratory tractinfections. Bronchoalveolar lavage (BAL) to collect respiratory tractfluid and tissue has become a standard method of ascertaining bothextracellular and intracellular antibiotic concentrations after systemicadministration of the antibiotic. Extracellular concentrations arecalculated from fluid reflecting ELF, and intracellular concentrationsare measured in ACs, including macrophages.

This study was designed to determine intra-pulmonary PK of Compound 1 inhealthy subjects; currently there is no information on the concentrationof Compound 1 in pulmonary compartments in humans.

Compound 1 has been shown to have in vitro activity against the mostcommon typical and atypical causes of community-acquired bacterialpneumonia (CABP) and is being developed for the treatment of CABP. It isvery active in vitro against most Gram-positive pathogens. It alsoexhibits activity against atypical pathogens (e.g., Legionella species(spp.), Chlamydophila spp.), and some anaerobic and Gram-negativepathogens. The drug is active against strains expressing both mechanismsof tetracycline resistance, as well as strains that are resistant tocurrently available antibiotics, including methicillin, vancomycin,erythromycin, and ciprofloxacin. The in vitro activity of Compound 1 wasnot affected by serum or lung surfactant, an important characteristicthat is consistent with potential utility in infections involving thelower respiratory tract. Further, it has been shown to be effective inmouse models of lower respiratory tract infections caused byStreptococcus pneumoniae and Haemophilus influenzae. In mice, Compound 1concentrations in lung tissue exceed plasma concentrations by 3.7 to 4.4fold. In vitro results against intracellular bacteria and tissue cultureexperiments indicate that Compound 1 concentrates within mammaliancells.

Tigecycline has a similar PK profile to Compound 1, and with itsdocumented concentration levels achieved in human ELF, the inclusion oftigecycline is intended to provide assay sensitivity in the study.

Compound 1 has been developed for both iv and oral administration andhas been well characterized in 16 Phase 1 studies including 536 subjectsexposed to Compound 1. In addition, Compound 1 was evaluated in a Phase2 study of 219 subjects with complicated skin and skin structureinfection (cSSSI) and a sponsor-terminated Phase 3 study that enrolled143 subjects with cSSSI. Compound 1 was well-tolerated and demonstratedefficacy similar to an established comparator (linezolid).

The purpose of the study was to determine the concentration of Compound1 in pulmonary compartments (ELF and in pulmonary ACs, including AMs)and define time course of pulmonary distribution in comparison with theplasma pharmacokinetic (PK) profile. Compound 1 was administeredintravenously to healthy subjects for 4 days (5 doses) to steady-statesystemic concentrations. Subjects were grouped to have BAL at set timesafter the last dose of drug. Concomitant blood samples were taken tocompare tissue with serum antibiotic concentrations.

The data obtained in this study, namely the in vitro activity againstcommon typical and atypical pathogens and the sustained ELF and AC(mostly AM) concentrations for 24 hours suggest that Compound 1 has thepotential to be a useful antibacterial agent for the treatment of lowerrespiratory tract bacterial infections caused by susceptible pathogens.

Tigecycline has a similar PK profile to Compound 1, and with its abilityto achieve concentrations in ELF, the inclusion of tigecycline providedassay sensitivity in the study.

Thus the primary objective of this study was to determine concentrationsof Compound 1 in ELF and AC (mostly AM) and define the time course ofpulmonary distribution with concurrent plasma PK sampling of Compound 1in healthy adult subjects.

The secondary objective was to evaluate the PK of Compound 1 inpulmonary and plasma compartments in healthy adult subjects.

The exploratory objective was to determine the PK of Compound 1 inpulmonary and plasma compartments compared to tigecycline PK inpulmonary and plasma compartments in healthy adult subjects.

This study was designed as a single-center, multiple-dose, open-labelstudy to determine concentrations of Compound 1 and tigecycline inpulmonary compartments (ELF and AC) in healthy adult subjects afteradministration of Compound 1 and tigecycline to steady state levels ofdosing. Enrollment of approximately 62 subjects was planned to ensureapproximately 42 subjects receive Compound 1 and to have abronchoalveolar lavage (BAL) performed and approximately 20 subjectsreceived tigecycline and had a BAL performed. Additional subjects wereto be enrolled to replace subjects who discontinued or were screenfailures.

Approximately 42 subjects received 5 doses of Compound 1 100 mgintravenously (30 minute infusions at t=0, 12, 24, 48, and 72 hours);Compound 1 subjects were assigned equally to 1 of 7 BAL sampling timepoints (approximately 6 subjects per BAL time point group) forcollection of ELF and AC samples.

Upon completion of the study, 42 enrolled subjects received Compound 1(69% male, median age 36 y, median BMI 27 kg/m²). Six subjects had BALat each of the 7 time points. One subject had a BAL sampling error andwas not included in BAL analyses.

Compound 1 concentrations in ELF and AC were measured at only 1observation time for each subject, and data from all subjects werepooled for a PK analysis of the mean intrapulmonary concentrations. Theconcentration of Compound 1 in BAL fluid was normalized by the dilutionfactor in respect to urea levels detected in plasma and BAL.

One standard bronchoscopy was performed for each subject. Within 1 BALtimepoint group, all subjects were assessed at the same timepoint,either at 0.5, 1, 2, 4, 8, 12, and 24 hours after the last doseadministration on Day 4. A blood sample was taken at the time ofbronschoscopy for plasma urea measurement. Additionally, blood sampleswere collected for plasma PK assessment at 0, 0.5 (end of infusion), 1,1.5, 2, 3, 4, 6, 8, 12, and 24 hours post-dose for all subjects on Day4.

Blood samples were taken and BAL performed in Compound 1-treatedsubjects according to the schedule below:

Day 1 Day 2 Day 3 Day 4 Compound 1 Dose 1 Dose 2 Dose 3 Dose 4 Dose 5100 mg iv t = 0 h t = 12 h t = 24 h t = 48 h t = 72 h Blood sample forPre-dose N/A N/A N/A Predose, 0.5 (end of infusion), plasma PK(baseline) 1, 1.5, 2, 3, 4, 6, 8, 12, and (all subjects) 24 h Takenright before BAL (±3 min), from the arm contralateral to the infusionsite BAL sample N/A N/A N/A N/A Subjects assigned to either 0.5(approximately (end of infusion), 1, 2, 4, 8, 6 subjects for each time12, or 24 h point) Blood sample for N/A N/A N/A N/A 0.5 (end ofinfusion) 1, 2, 4, 8, plasma urea 12, or 24 h, taken right beforemeasurement (same BAL (±3 min) time point as BAL; 1 sample per subject)BAL = bronchoalveolar lavage; h = hour; N/A = not applicable; PK =pharmacokinetic; t = time.

For the patients receiving Compound 1, Mean (±SD) plasma pharmacokineticparameters after the fifth Compound 1 dose included maximumconcentration of 2.26±0.76 μg/mL, volume of distribution of 165±58 L,clearance of 8.03±1.43 L/h, and elimination half-life of 14.7±4.2 h.Mean (±SD) Compound 1 concentrations (μg/mL) at time of bronchoscopy andBAL were:

Sampling Time Plasma ELF AM 0.5 h 1.80 ± 0.13 1.73 ± 1.01 14.26 ± 9.30 1h 0.89 ± 0.19 2.25 ± 0.72 12.80 ± 8.48 2 h 0.93 ± 0.33 1.51 ± 0.94 10.77± 7.59 4 h 0.59 ± 0.15 0.95 ± 0.33 17.99 ± 7.17 8 h 0.56 ± 0.12 0.58 ±0.19 12.27 ± 4.70 12 h 0.42 ± 0.07 0.61 ± 0.29 12.29 ± 4.61 24 h 0.27 ±0.05 0.41 ± 0.13 10.36 ± 4.04

Penetration ratios based on AUC₀₋₂₄ values of mean and median ELF andplasma concentrations were 1.47 and 1.42, whereas ratios of AC (AlveolarCell, mostly AM) to plasma concentrations were 25.8 and 24.8.

Approximately 20 subjects received 1 dose of 100 mg tigecyclineintravenously (30 minute infusion at t=0) followed by 6 doses of 50 mgtigecycline intravenously (30 minute infusions at t=12, 24, 36, 48, 60,and 72 hours). Tigecycline subjects were assigned equally to 1 of 4 BALsampling timepoints (approximately 5 subjects per BAL timepoint group)for collection of ELF and AC samples. Tigecycline concentrations in ELFand AC were measured at only 1 observation time for each subject, anddata from all subjects were pooled for a PK analysis of the meanintrapulmonary concentrations. The concentration of tigecycline in BALfluid was normalized by the dilution factor in respect to urea levelsdetected in plasma and BAL.

Blood samples were taken and BAL performed in tigecycline-treatedsubjects according to the schedule below:

Day 1 Day 2 Day 3 Day 4 Tigecycline Dose 1 Dose 2 Dose 3 Dose 4 Dose 5Dose 6 Dose 7 iv 100 mg 50 mg 50 mg 50 mg 50 mg 50 mg 50 mg t = 0 h t=12 h t = 24 h t = 36 h t = 48 h t = 60 h t = 72 h Blood sample forPre-dose N/A N/A N/A N/A N/A Predose, 0.5 (end of plasma PK (base line)infusion), 1, 1.5, 2, (all subjects) 3, 4, 6, 8, and 12 h Taken rightbefore BAL (±3 min), from the arm contralateral to the infusion site BALsample N/A N/A N/A N/A N/A N/A Subjects assigned to (approximately 5either 2, 4, 6, or 12 h subjects for each timepoint) Blood sample forN/A N/A N/A N/A N/A N/A 2, 4, 6, or 12 h, plasma urea taken right beforemeasurement (same BAL (±3 min) timepoint as BAL; 1 sample per subject)BAL = bronchoalveolar lavage; h = hour; N/A = not applicable; PK =pharmacokinetic; t = time.

Safety assessments included physical examinations, electrocardiograms(ECG), vital signs, standard clinical laboratory evaluations (bloodchemistry, hematology), pregnancy testing, and adverse event (AE) andserious adverse event (SAE) monitoring.

Treatment-emergent adverse events (TEAEs) were reported in 29%. The mostcommon TEAE was headache (12%). There were no severe or serious TEAEs,and no discontinuations due to TEAEs in Compound 1 treated subjects.There were no clinically significant changes in vital signs, laboratoryor ECG parameters.

The duration of treatment was designed to ensure that both treatmentarms were at steady state at the time of BAL.

During the bronchoscopy 4 aliquots of 50 mL each of sterile salinesolution were instilled into the right lobe of the lungs, immediatelyaspirated, and placed on ice. The first bronchoalveolar lavage aliquotinstilled (BALX) fraction was prepared and analyzed separately fromsubsequent instillations. The aspirate from the second through thefourth instillation, once pooled, represented the BAL fraction. Analiquot of BAL was removed and used to determine the cell count anddifferential cell composition including pulmonary macrophages. Theremaining supernatant was centrifuged and the supernatant and cellpellet were immediately frozen pending analysis. An aliquot of the BALsupernatant was frozen separately for urea assay. The amount of testarticle detected in the cell pellet was normalized by the correspondingfraction of macrophages. Fraction BALX was analyzed in a similar manner,except the cell pellet was not assessed or assayed. A blood sample todetermine urea concentration was obtained at the time of the secondinstillation (±3 minutes). The concentration of test article detected inthe BALX and BAL fluid was normalized by the dilution factor withrespect to urea levels detected in the BALX and BAL fractionsrespectively.

Subjects participated in the study for approximately 34 days. Followingscreening, eligible subjects were randomly assigned to Compound 1 ortigecycline treatment and assigned to a BAL lavage sample timepoint.Subjects had Baseline evaluations on Day −1 and then received testarticle treatment for 4 days followed by BAL at their assignedtimepoint. A Study Completion visit was conducted on the day followingthe final test article dose. There was a Final Follow-up assessment 7 to14 days following the subject's last dose of test article which may becompleted by telephone contact or other interactive technology unless anexamination was needed to evaluate AEs or abnormalities noted at theStudy Completion visit.

Safety assessments included physical examinations, electrocardiograms(ECG), vital signs, standard clinical laboratory evaluations (bloodchemistry, hematology), pregnancy testing, and AE and SAE monitoring.

Dosing Regimen:

The current intended therapeutic dose of Compound 1 for the treatment ofCABP is 100 mg iv every 12 hours (q12h) for 2 doses followed by every 24hours (q24h), with the option to switch to 300 mg orally q24h.

The approved therapeutic dose of tigecycline (TYGACIL®) for thetreatment of CABP is 100 mg iv for 1 dose followed by 50 mg iv q12h.

Patient Inclusion/Exclusion Criteria

Enrolled patients were male or female subjects age 18 to 55 years ofage, in good health as determined by past medical history, physicalexamination, vital signs, ECG, and laboratory tests (no clinicallysignificant abnormalities in the opinion of the investigator). Vitalsigns (oral body temperature, systolic and diastolic blood pressure(BP), and pulse rate) were assessed in a sitting position after thesubject had rested for at least 3 minutes. Sitting vital signs should bewithin the following ranges: oral body temperature, 35.0° C. to 37.5° C.(95.0° F. to 99.5° F.); systolic BP, 90 to 140 mm Hg; diastolic BP, 50to 90 mm Hg; pulse rate, 40 to 90 bpm; Blood pressure and pulse wereassessed again after 3 minutes in a standing position at the Screeningvisit. There were no more than a 20 mm Hg drop in systolic or 10 mm Hgdrop in diastolic BP and increase in heart rate (>20 bpm) associatedwith clinical manifestation of postural hypotension. Out-of-range vitalsigns may be repeated once at the discretion of the investigator, ifnecessary.

Subjects weighed at least 50 kg, and had a body mass index (BMI) withinthe range of ≥18.0 to ≤30.0 kg/m². Females had a negative serumpregnancy test at the Screening and Baseline visits and agreed to complywith using an acceptable form of birth control from Screening throughthe final follow-up assessment. Males agreed to use an acceptable methodof birth control with female partner(s) and did not donate sperm fromScreening through the Final Follow-up assessment.

Patients were excluded from the study if one or more of the followingwas present:

-   -   Use of other investigational drugs within 5 half-lives or 30        days prior to Screening, whichever is longer.    -   Had a history of hypersensitivity or allergic reaction (e.g.,        anaphylaxis, urticaria, other significant reaction) to any        tetracycline (e.g., minocycline, doxycycline, or tigecycline).    -   Had a history of clinically significant ECG abnormalities, or        any of the following ECG abnormalities at the Screening or        Baseline visit: PR>220 msec, QRS complex >120 msec, Long QT        syndrome, QTc Fridericia's Correction Formula (QTcF) >450 msec        (males), and QTcF >470 msec (females)    -   Pregnant or nursing (breastfeeding) women.    -   History of malignancy of any organ system (other than localized        basal cell carcinoma of the skin), treated or untreated, within        the past 5 years, regardless of whether there is evidence of        local recurrence or metastases.    -   Use of tobacco products in the 3 months prior to Screening.    -   A positive urine cotinine test at Screening or Baseline.    -   Use of any prescription drugs or herbal supplements within 4        weeks prior to the Baseline visit, and/or over-the-counter (OTC)        medications including dietary and fitness/body-building        supplements (vitamins included) within 2 weeks prior to the        Baseline visit.    -   Donation or loss of 400 mL or more of blood or plasma within 8        weeks prior to the Baseline visit, or longer if required by        local regulation.    -   Hemoglobin levels <12.5 g/dL for males, <11.5 g/dL for females        at Screening/Baseline visit.    -   Significant illness within 2 weeks prior to Baseline visit.    -   History of autonomic dysfunction (e.g., recurrent episodes of        fainting, palpitations, etc) within 3 years prior to Screening.    -   History of acute or chronic bronchospastic disease (including        asthma and chronic obstructive pulmonary disease (COPD), treated        or not treated) within 3 years prior to Screening.    -   Any surgical or medical condition which, in the opinion of the        investigator, might significantly alter the absorption,        distribution, metabolism, or excretion of drugs, or which may        jeopardize the subject in case of participation in the study.    -   History of, or active, inflammatory bowel disease, ulcers, GI or        rectal bleeding, or pancreatitis.    -   Liver disease or liver injury as indicated by abnormal LFTs such        as aspartate aminotransferase (AST), ALT, gamma-glutamyl        transpeptidase (GGT), CK, alkaline phosphatase (ALP), or serum        bilirubin. The investigator should be guided by the following        criteria: Serum bilirubin may not exceed 1.2 times the upper        limit of normal (ULN); Any other single parameter listed above        may not exceed 1.5 times ULN; any elevation of more than 1        parameter excludes a subject from participation in the study.        Testing may be repeated once more as soon as possible to rule        out lab error. Recheck results must not meet the criteria above        in order for subject to qualify.    -   History or presence of impaired renal function as indicated by        clinically significant abnormal creatinine or blood urea        nitrogen (BUN) and/or urea values, or abnormal urinary        constituents (e.g., albuminuria).    -   Evidence of urinary obstruction or difficulty in voiding at        Screening.    -   Was known to be human immunodeficiency virus (HIV)-positive.    -   Had known chronic hepatitis B or chronic hepatitis C infection.    -   Positive alcohol test or positive drug screen at the Screening        or Baseline visit.    -   Had previously been treated with Compound 1 or previously        enrolled in this study.    -   Any concomitant condition that, in the opinion of the        investigator, is likely to interfere with the determination of        AEs or completion of the study procedures.

Procedures

There were 3 protocol-defined phases of the study: Screening, Treatmentand Follow-up. The study had the following protocol-defined evaluations:

-   -   Screening visit    -   Baseline visit    -   Visits on Days 1, 2, and 3 (test article dosing) Visit on Day 4        (test article dosing and BAL)    -   Study Completion visit on Day 5    -   Final Follow-up assessment: Study Day 11-18 (7-14 days after the        last dose of test article)

Subjects who discontinued study treatment prematurely had the StudyCompletion procedures performed prior to clinic discharge and a FinalFollow-up assessment.

Screening was used to establish subject eligibility and Baselinecharacteristics for each subject. The following information wascollected: review of inclusion/exclusion criteria; relevantmedical/surgical history and current medical conditions (predisposingfactors that may affect lung function (e.g., prior lung infection, mildto moderate COPD, asthma, history of smoking, chronic cough, etc);demographics; physical examination; urine alcohol test, drug screen,cotinine test; vital signs; 12-lead ECG; laboratory tests (hematology,chemistry, coagulation, pregnancy test (women only)); and AEs since thesigning of the ICF Concomitant medications (past 4 weeks).

The treatment period was 5 days in duration. Subjects who met inclusioncriteria and did not meet exclusion criteria received their first doseof test article. The following assessments were done: Vital signs; AEs;Concomitant treatments; and Test article administration andaccountability.

During the Intravenous Treatment Phase (Test Article), infusions ofCompound 1 were administered continuously over approximately 30 minutes(at least 30 minutes and not more than 45 minutes) according to theschedule in Table 1-3 below. Infusions of tigecycline were administeredcontinuously over approximately 30 minutes (at least 30 minutes and notmore than 45 minutes) according the schedule in Table 1-4. All infusionstart and stop times and compliance (delivery of ≥90% of the dose) wererecorded in source documents and on the eCRF.

TABLE 1-1 Treatment Regimen for IV Compound 1 Time^(a) Dose^(b) t = 0 hCompound 1 100 mg in 100 mL NS t = 12 h Compound 1 100 mg in 100 mL NS t= 24 h Compound 1 100 mg in 100 mL NS t = 48 h Compound 1 100 mg in 100mL NS t = 72 h Compound 1 100 mg in 100 mL NS t = time; NS = normalsaline (0.9% sodium chloride) for injection ^(a)The start time of thestart of the first infusion was designated time 0 (t = 0 h). Subsequentinfusions were administered within ±1 hour of the target infusion timeslisted. ^(b)All 100 mL infusions of Compound 1 were administeredcontinuously over 30 min (at least 30 min and not more than 45 min).

TABLE 1-2 Treatment Regimen for IV Tigecycline Time^(a) Dose^(b)

TABLE 1-1 Treatment Regimen for IV Compound 1 Time^(a) Dose^(b) t = 0 htigecycline 100 mg in 100 mL NS t = 12 h tigecycline 50 mg in 100 mL NSt = 24 h tigecycline 50 mg in 100 mL NS t = 36 h tigecycline 50 mg in100 mL NS t = 48 h tigecycline 50 mg in 100 mL NS t = 60 h tigecycline50 mg in 100 mL NS t = 72 h tigecycline 50 mg in 100 mL NS t = time; NS= normal saline (0.9% sodium chloride) for injection. ^(a)The start timeof the start of the first infusion was designated time 0 (t = 0 h).Subsequent infusions were administered within ±1 hour of the targetinfusion times listed. ^(b)All 100 mL infusions of tigecycline wereadministered continuously over 30 min (at least 30 min and not more than45 min).

During the Follow-up Phase, subjects were evaluated at 2 visits afterthe completion of treatment: at the Study Completion visit on Day 5 andat a Final Follow-up assessment on Study Days 11 to 18 (7 to 14 daysafter subject's last dose of test article). The Final Follow-upAssessment was conducted by telephone contact unless an examination wasneeded to evaluate AEs or abnormalities noted at Study Completion.

For IV injection, Compound 1 was supplied as 100 mg (plus 4% overfill)sterile, lyophilized powder for reconstitution packaged in a clear,glass vial with a rubber stopper and aluminum overseal. Excipientsincluded tosylate acid counter ion, sucrose, hydrochloric acid andsodium hydroxide to adjust the pH. Each vial was reconstituted into aclear solution by adding 5 mL Sterile Water for Injection. The vial wasswirled gently to ensure complete dissolution prior to use. Excessiveshaking was avoided to prevent foaming. Reconstituted vials were usedimmediately to prepare the infusion solution. The infusion solution wasprepared by withdrawing 5 mL of reconstituted solution from the vial andslowly injecting into a 100 mL normal saline (NS) for injection (0.9%sodium chloride) infusion bag. The prepared infusion solution was usedwithin 8 h or stored at up to 24 h at 2° C. to 8° C. (35.6° F. to 46.4°F.). The 100 mL infusion solution was administered at room temperaturecontinuously over 30 minutes (at least 30 minutes and not more than 45minutes).

Reconstituted Compound 1 was injected as 100 mg iv q12h for first 2doses followed by 100 mg iv q24h (starting 24 h after first dose) for2-3 doses. Total treatment consisted of 5 doses, and a duration of 5days.

The comparator test article Tigecycline was injected as 100 mg iv firstdose followed by 50 mg iv q12h for 6 doses. Total treatment consisted of7 doses, for a duration of 5 days.

Safety

Any subject who received at least 1 dose of test article was included inthe evaluation for safety. Safety was assessed by the followingmeasures: Physical exams; AEs and SAEs; Vital signs; Laboratoryassessments (Blood samples for hematology, chemistry, and coagulation(prothrombin time only)); ECG; and Pregnancy assessments.

Data Analysis

All analyses of data for this study complied with InternationalConference on Harmonisation of Technical Requirements for Registrationof Pharmaceuticals for Human Use (ICH-E9) and the sponsor's guidancedocuments and standards. Statistical analyses were performed usingStatistical Analysis Software (SAS).

a) Analysis Populations

Subject populations have been defined for the various analyses of PKsand safety, as follows:

-   -   The PK population consisted of all subjects who received test        article and had at least 1 evaluable PK parameter.    -   The Safety population consisted of all subjects who received at        least 1 dose of test article.

b) Subject Demographics and Baseline Characteristics

Descriptive statistics, by treatment arm, was provided for thefollowing: Subject disposition (completed test article, discontinuedtest article by reason for discontinuation, completed study,discontinued study by reason for discontinuation); Protocol deviations;Medical histories and continuing medical conditions.

Baseline demographic and medical variables were analyzed using a 2-sidedFisher's exact test (for categorical variables) or a 2-sided WilcoxonRank Sum test (for ordinal and continuous variables).

c) Safety Outcome Measures

Safety variables included the incidence rate of AEs, change in vitalsigns, ECG parameters, and laboratory test results obtained during thecourse of the study. Subjects were analyzed according to the treatmentactually received.

d) PK

All completed subjects with evaluable plasma PK parameter data wereincluded in the PK data analysis. The following Compound 1 andtigecycline plasma PK parameters were determined:

-   -   Area under the curve (AUC) from time 0 to 24 hours after dosing        (AUC₀₋₂₄) for Compound 1, and AUC from time 0 to 12 hours after        dosing (AUC₀₋₁₂) for tigecycline,    -   Maximum plasma concentration (C_(max)),    -   Time to maximum plasma concentration (T_(max)) and    -   Terminal elimination half-life (T_(1/2)) associated with the        terminal slope of the semilogarithmic concentration-time curve.

All the biofluid concentrations were expressed in ng/mL. Allconcentrations below the limit of quantification (BLQ) or missing datawere labeled as such in the concentration data listings. ConcentrationsBLQ were treated as zero in summary statistics for concentration dataonly. They were not considered for calculation of PK Parameters (withthe exception of the predose samples). PK parameters were determinedusing non-compartmental method(s) using WinNonlin Pro.

The primary objective of the study was to assess the concentration ofCompound 1 in lung ELF and AC (mostly AM). The concentration of Compound1 and tigecycline in lung ELF and AC were calculated as area under the(concentration/time) curve in epithelial lung fluid (AUC_(ELF))/areaunder the (concentration/time) curve in plasma (AUC_(plasma)) and areaunder the (concentration/time) curve in alveolar cells(AUC_(AC))/AUC_(plasma), respectively. These were obtained from theabove plasma PK parameter calculations (AUC₀₋₂₄ and AUC₀₋₁₂). Thetigecycline concentration information was serve as an internalvalidation for assay sensitivity purposes.

To determine the AUC_(ELF) parameters for Compound 1 and tigecycline,their concentration data were used by deriving from the correspondingBAL concentrations. The concentration of Compound 1 or tigecycline inthe lung ELF (C_(ELF)) was calculated as:C_(ELF)=C_(BAL)*(V_(BAL)/V_(ELF)), where C_(BAL) is the concentration inthe BAL fluid, V_(BAL) is the volume of the aspirated BAL fluid, andV_(ELF) is the volume of lung ELF.

The volume of the lung ELF within the BAL fluid was estimated by(urea_(BAL)/urea_(plasma)), where urea_(BAL) and urea_(plasma)represented the concentrations of urea in the BAL fluid and plasma,respectively.

Compound 1 and tigecycline concentration data from ELF, AC, and AM werepresented in a listing. AUC_(ELF) and AUC_(AC) were determined frompooled pulmonary concentration data and were summarized by treatmentgroup. For the Compound 1 cohort there were at least 6 ELF and ACconcentrations for each of the 7 timepoints over the 24 hour dosinginterval. For the tigecycline cohort there were 5 ELF and ACconcentrations for each of the 4 timepoints over the 12 hour dosinginterval.

The calculated concentration of Compound 1 and tigecycline in lung ELFand AC as the ratios of AUC_(ELF)/AUC_(plasma) andAUC_(AC)/AUC_(plasma), were summarized by treatment group. BALX wasassessed separately and in an identical fashion. The impact of “BALX” onthe PK understanding of Compound 1 and tigecycline was assessed.

PK Samples for Compound 1 or Tigecycline Concentration

The permitted windows for PK sample collection were as follows:

Window PK blood collection- scheduled time relative to test article dosePredose within 10 min before test article administration 0.5 to 1 h  ±2min 1.5 h to 6 h  ±5 min 8 h to 24 h ±15 min Sample collected at thebronchoscopy ±3 min of the second BAL instillation scheduled timeBronchoscopy - BAL and AC collection Scheduled time ±15 min Blood samplefor plasma urea testing Sample collected at the bronchoscopy ±3 min ofthe second scheduled time BAL instillation

For PK blood collection and processing, all blood samples were taken byeither direct venipuncture or an indwelling cannula inserted in aforearm vein (on the arm contralateral to the infusion site) at thetimepoints specified in the Study Flowchart. The PK blood samplecollected at the bronchoscopy scheduled time was obtained at the time ofthe second BAL instillation (±3 minutes) from the arm contralateral tothe infusion site.

Blood samples were collected into labeled 4 mL tubes containing sodiumheparin. Immediately after the sample was collected, the tube was gentlyinverted 5 to 8 times to thoroughly mix the anticoagulant and thenplaced upright in a cryoblock or test tube rack surrounded by ice untilcentrifugation. Samples were centrifuged at 1500×g (gravity) for 10minutes at approximately 4° C. within 30 minutes of collection. Theresultant plasma was divided into 2 equal aliquots, placed in individualcryovials, and immediately frozen at −70° C. or colder within 1 hour ofcollection. The tubes were kept frozen at −70° C. or colder pendingshipment to the bioanalytical laboratory.

For BAL and AC sample collection and handling, one standard bronchoscopywas performed in each subject following the last administration of testarticle at the timepoints specified in the Study Flowchart. Subjectswere continuously monitored during bronchoscopy. Blood pressure, heartrate, and respiratory rate were recorded just prior to (within 30minutes) the bronchoscopy procedure scheduled time, 30 minutes after,and 60 minutes after of the bronchoscopy procedure scheduled time.Topical lidocaine, 4% solution for the oropharynx, and 2% solution forthe nasopharynx, were applied to the upper airways to prepare subjectsfor bronchoscopy. If needed, a 1% lidocaine solution was used in thelower airways. A fiberoptic bronchoscope was inserted into the middlelobe of the right lung. Four-50 mL aliquots of sterile 0.9% salinesolution were instilled into the right lobe and immediately aspirated,and placed on ice. The first 50 mL instilled (fraction BALX) wascollected (2 aliquots of 4 mL), immediately placed on ice, the volumerecorded, stored, prepared, and analyzed separately from subsequentinstillations. The aspirate from the second through the fourthinstillation was collected immediately, placed on ice, and the volumerecorded. The collective aspirates (second through fourth), once pooled,represented the BAL fraction.

An aliquot of BAL was removed and used to determine the cell count anddifferential cell composition. The fraction of pulmonary macrophages wasdetermined during cell count and differentiation. An aliquot of the BALXand BAL supernatants were reserved for a urea assay. The remainingsupernatant was centrifuged immediately at 400×g for 5 minutes in arefrigerated centrifuge. Supernatant, fractionated into 5 mL aliquots,and the cell pellet were immediately frozen at −70° C. or colder pendinganalysis. Fraction BALX was analyzed in a similar manner, with theexception that the cell pellet was not assessed or assayed.

For urea assay in BAL fluid and plasma, a blood sample to determine ureaconcentration was obtained at the time of the second BAL instillation(±3 minutes). Urea blood samples were taken by either directvenipuncture or an indwelling cannula inserted in a forearm vein (on thearm contralateral to the infusion site). Plasma was prepared from theurea blood sample in the same manner as plasma samples for PK analysis.

Plasma samples, BAL supernatant samples and BALX supernatant sampleswere analyzed for urea concentration by the bioanalytical laboratory.

After all of the PK samples from a single subject had been collected andfrozen at −70° C. or colder, the primary samples from each timepointwere batched together with corresponding primary samples from othersubjects and be carefully packaged and shipped frozen at −70° C. orcolder to the bioanalytical laboratory designated by the sponsor.Samples were shipped with sufficient dry ice to remain frozen duringovernight transit. For each subject and timepoint, the remaining storedaliquots were retained on-site at −70° C. or colder until released orrequested by the sponsor.

The bioanalytical laboratory assayed the samples for Compound 1 ortigecycline using a specific, sensitive, and validated liquidchromatography/tandem mass spectrometry (LC/MS/MS) method approved bythe sponsor.

To determine ELF volume and concentrations of Compound 1 or Tigecyclinein ELF, urea measured in the BALX and BAL supernatant and in plasma wasused to calculate the volume of ELF using the conventional urea dilutionmethod. The volume of ELF was determined by the following equation:

V _(ELF) =V _(BAL)*(urea_(BAL)/urea_(plasma)),

where V_(ELF)=volume of ELF in the BAL sample, V_(BAL)=volume ofaspirated BAL fluid, urea_(BAL)=concentration of urea in BALsupernatant, and urea_(plasma)=concentration of urea in plasma.

The concentration of Compound 1 or tigecycline in ELF (C_(ELE)) wascalculated as:

C _(ELF) =C _(BAL)*(V _(BAL) /V _(ELF)),

where C_(ELF)=concentration of Compound 1 or tigecycline in ELF,C_(BAL)=measured concentration of Compound 1 or tigecycline in BALsupernatant, V_(ELF)=volume of ELF in the BAL sample, and V_(BAL)=volumeof aspirated BAL fluid.

To determine AC volume and concentration of Compound 1 or Tigecycline inAC and AM, the volume of AC collected in the BAL cell pellet suspensionwas determined from the BAL fluid cell count. The volume of cells wascalculated by multiplication of cell counts in ELF with the known volumeof ACs. The volume of cells was determined by multiplying the cell countwith the mean macrophage cell volume of 2.42 μL/10⁶ cell.

The measured concentration of Compound 1 or tigecycline in AC (CAC) wasdetermined by the following equation:

CAC=(C _(pellet suspension) /V _(AC)),

where C_(pellet suspension) is the Compound 1 or tigecyclineconcentration in 1 mL cell suspension and V_(AC) is the volume of AC in1 mL cell suspension.

The measured concentration of Compound 1 or tigecycline in AMs (C_(AM))was derived from CAC by adjusting for the percentage of macrophages andmonocytes in the AC as determined by a differential cell count of theBAL fluid.

FIG. 1 shows the result of mean Compound 1 concentration vs. timeprofile in AC, plasma, and ELF. It is apparent that the mean Compound 1concentration in AC (mostly AM) is at least an order of magnitude higher(about 25 fold) than that in plasma, while the mean Compound 1concentration in ELF is at least about 40% higher than that in plasma.See tables.

Sampling Time ELF to Plasma (Mean, SD) AM to Plasma (Mean, SD)0.5-hour^(a) 0.95 ± 0.56 8.12 ± 5.95 1-hour^(a) 2.72 ± 1.26 13.85 ±7.38  2-hour^(b) 1.50 ± 0.61 12.29 ± 6.57  4-hour^(a) 1.79 ± 0.49 34.72± 15.14 8-hour^(a) 1.07 ± 0.45 23.12 ± 11.17 12-hour^(a) 1.44 ± 0.5728.97 ± 8.78  24-hour^(a) 1.65 ± 0.86 40.33 ± 10.29 ^(a)6 reportedplasma, ELF, and AM concentrations at this sampling time ^(b)5 samplesplasma, ELF, and AM concentrations at this sampling time

Compound 1 Demonstrated Higher Epithelial Lining Fluid (ELF) andAlveolar Cellular (AC) Concentrations

Plasma ELF AC (μg*h/ (μg*h/ Ratio (μg*h/ Ratio mL) mL) ELF:Plasma mL)AC:Plasma AUC_(mean) 11.73 17.23 1.47 302.5 25.8 AUC_(median) 11.8016.74 1.42 292.3 24.8 AUC_(mean) = Area-under-the-curve based on meanconcentration at each BAL sampling time AUC_(median) =Area-under-the-curve based on median concentration at each BAL samplingtime

This data provides important information on the time course andmagnitude of extracellular and intracellular concentrations of Compound1 in the lung. Intravenous administration of Compound 1 at 100 mg as a30-minute infusion produced concentrations that were higher inepithelial lining fluid (ELF) and alveolar cellular (AC, including AM)than simultaneous plasma concentrations throughout the 24-hour periodafter five doses. The in vitro activity against common typical andatypical pathogens and the sustained ELF and AC/AM concentrations for 24hours suggest that Compound 1 can be a useful antibacterial agent forthe treatment of lower respiratory tract bacterial infections caused bysusceptible pathogens.

Safety Monitoring

An AE is any untoward, undesired, or unplanned event in the form ofsigns, symptoms, disease, or laboratory or physiologic observationsoccurring in a person given a test article or in a clinical study. Theevent does not need to be causally related to the test article orclinical study. An AE includes, but is not limited to, the following:any clinically significant worsening of a preexisting condition; an AEoccurring from overdose of a test article, whether accidental orintentional (overdose is a dose greater than that specified in theprotocol); an AE occurring from abuse (e.g., use for nonclinicalreasons) of a test article; and an AE that has been associated with thediscontinuation of the use of a test article.

A SAE is an AE that: results in death; is life-threatening; requireshospitalization or prolongation of an existing hospitalization; resultsin a persistent or significant disability or incapacity; results in acongenital anomaly or birth defect; or additionally, important medicalevents that may not result in death, be life-threatening, or requirehospitalization may be considered SAEs when, based on appropriatemedical judgment, they may jeopardize the subject and may requiremedical or surgical intervention to prevent 1 of the outcomes listed inthis definition. Examples of such events include allergic bronchospasmrequiring intensive treatment in an emergency room or at home, blooddyscrasias or convulsions that do not result in hospitalization, ordevelopment of drug dependency or drug abuse.

If there was any doubt about whether the information constitutes an SAE,the information was treated as an SAE.

A protocol-related AE is an AE occurring during a clinical study that isnot related to the test article, but is considered by the investigatoror the Medical Monitor (or designee) to be related to the researchconditions, i.e., related to the fact that a subject is participating inthe study. For example, a protocol-related AE may be an untoward eventrelated to a medical procedure required by the protocol.

Example 2 Effect of Food on the Bioavailability of Omadacycline inHealthy Volunteers

Compound 1 (9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline) is afirst-in-class aminomethylcycline antibiotic that is characterized byimproved in vitro antimicrobial activity (Honeyman et al, AntimicrobAgents Chemother. 59:7044-7053, 2015).

Phase 3 studies have concluded with Compound 1 as oral and intravenous(IV) monotherapy in patients with acute bacterial skin and skinstructure infection (ABSSSI). During the development process, oralCompound 1 formulations have evolved from free-base in a capsule througha series of tablet and salt formulations in order to optimize oralbioavailability while improving tolerability. The current phase 3 tabletformulation is the tosylate salt of Compound 1, which has been shown tohave an absolute bioavailability of 34.5% when administered underfasting conditions. The primary objective of this study was to evaluatethe relative bioavailability of a single oral 300 mg dose of Compound 1(administered as the phase 3 tablet formulation) at various times afterthe consumption of food in healthy adult subjects.

The result of this study showed that food consumption has an effect onthe oral bioavailability of a single 300 mg Compound 1 dose.

Briefly, the study was a phase 1, randomized, open-label 4-period,crossover study. Before dosing on Day 1 of Period 1, subjects wererandomized to one of four treatment sequences (see Table 2-1). On Day 1of each period, subjects received a single oral dose of 300 mg Compound1 (2×150 mg tablets) at various times after the consumption of food.There was a washout period of at least 5 days between each dosingperiod. A final study completion visit occurred 6 to 10 days after thelast dose of Compound 1.

TABLE 2-1 Treatment Sequences Sequence Period 1 Period 2 Period 3 Period4 ADBC A D B C BACD B A C D CBDA C B D A DCAB D C A B A: subjects fastedovernight (no food or drink except for water for at least 6 hours beforedosing); a standard high-fat (nondairy) meal was served 3 hours afterdosing B: a standard high-fat (nondairy) meal completed at 4 hoursbefore dosing C: a standard high-fat (nondairy) meal completed at 2hours before dosing D: a standard high-fat meal including dairycompleted at 2 hours before dosing

The high-fat (approximately 50% of total caloric content of the meal)and high-calorie (approximately 800 to 1000 calories) meal followed Foodand Drug Administration guidance recommendations, and providedapproximately 150, 250, and 500 to 600 calories from protein,carbohydrate, and fat, respectively (FDA Guidance, 2002). These mealswere to be consumed within 20 minutes. Dose administration forTreatments B, C, and D was based off of the end time of the meal. Duringall 4 treatment periods, subjects received no food or drink except waterfor at least 3 hours after dosing and no dairy products, antacids ormultivitamins for 4 hours after dosing.

A total of 32 subjects were enrolled and dosed in at least one treatmentperiod. Overall mean age was 32.3 years, with a range of 21 to 50 years;47% were male (Table 2-2). One subject was discontinued from the studybecause of a positive alcohol screen at baseline of period 3 and did notreceive Treatments A and D. One subject requested withdrawal and did notreceive Treatments B and C. PK data were available for 31 subjects foreach treatment condition.

TABLE 2-2 Baseline Demographics Subjects (n = 32) Age, years^(a) 32.2(8.0) Age range, years 21-50 Male, n (%) 15 (46.9) Race, n (%) white 24(75) black/African American 8 (25) Hispanic/Latino 12 (37.5) Height,cm^(a) 168.0 (9.5) Weight, kg^(a) 71.5 (13.4) BMI (body Mass Index),kg/m^(2a) 25.2 (3.2) ^(a)Mean (Standard Deviation)

Blood samples for pharmacokinetic (PK) assessments of Compound 1 werecollected before dosing (predose) and at 0.5, 1, 1.5, 2, 2.5, 3, 4, 6,8, 12, 16, and 24 hours after dosing in each period. PK parametersincluded: Area under the plasma concentration-time curve (AUC) from time0 to 24 hours after dosing (AUC₀₋₂₄); AUC from time 0 to the lastquantifiable concentration (AUC₀-t); AUC time 0 extrapolated to infinity(AUC_(0-inf)); Maximum (peak) observed plasma concentration (C_(max));Time to reach C_(max) (T_(max)); Terminal elimination half-life(T_(1/2)); Terminal phase rate constant (λz).

Safety and tolerability was assessed by: Adverse events (AEs); vitalsign measurements at multiple time-points within 24 hours post-dose ineach treatment period; and clinical laboratory tests 24 hours post-dosein each treatment period.

For statistical analysis, individual PK parameters for Compound 1 weresummarized with descriptive statistics. Geometric means were determinedfor AUC and C_(max). PK parameters were evaluated using noncompartmentalanalysis using Phoenix® WinNonlin® (Pharsight Corp, St. Louis, Mo.),Version 6.2.1. Confidence intervals (CI) for test treatments (fedstates: Treatments B, C, and D) compared with the reference treatment(fasted state: Treatment A) were constructed for AUC₀₋₂₄, AUC_(0-t),AUC_(0-inf), and C_(max). Absence of the effect of food was concluded ifthe 90% CI for the test-to-reference ratios (B/A, C/A, or D/A) ofgeometric means were contained within the criterion interval of 80% to125% for AUC₀₋₂₄, AUC_(0-t), AUC_(0-inf), and C_(max). For T_(max), theWilcoxon signed rank test was performed. p≤0.05 was consideredstatistically significant.

A linear mixed-effect model with treatment condition, sequence, andperiod as fixed effects and subject nested within sequence as a randomeffect was fitted to the natural log-transformed PK parameters forestimation of effects and 90% confidence intervals (CIs) for the fedstates compared with the fasted state.

TABLE 2-3 Plasma PK Parameters for Compound 1 after a Single 300 mg OralDose Mean (Coefficient of Variation) Treatment A Treatment B Treatment CTreatment D Parameter N = 31 N = 31 N = 31 N = 31 AUC₀₋₂₄, mcg*h/mL 7.2(28.1) 6.1 (26.3) 4.2 (23.4) 2.8 (44.3) AUC_(0-t), mcg*h/mL 7.2 (28.1)6.1 (26.3) 4.2 (23.4) 2.8 (44.3) AUC_(0-inf), mcg*h/mL 10.2 (27.0)^(b)8.8 (29.2) 6.0 (25.4) 4.0 (44.1) C_(max), mcg/mL 0.6 (25.3) 0.6 (25.0)0.4 (22.4) 0.3 (42.6) T_(max), h^(a) 2.5 (1.5, 4.1) 2.9 (1.0, 6.9) 2.9(1.0, 6.0) 2.9 (1.0, 6.0) T_(1/2), h 13.8 (10.3)^(b) 13.6 (12.7) 13.6(12.2) 13.5 (14.7) PK analysis showed that, for the 31 subjects includedin the PK analysis, fasted AUC_(0-inf), AUC_(0-t), and AUC₀₋₂₄ were10.2, 7.2, and 7.2 mcg*h/mL, respectively, and C_(max) was 0.6 mcg/mL.Across all treatment periods, mean T_(1/2) ranged from 13.5 to 13.8hours, and median T_(max) ranged from 2.5 to 2.9 hours. Notreatment-related adverse events or clinically relevant changes inlaboratory values, or vital signs occurred. See Table 2-3.

A significant reduction in systemic exposure to omadacycline wasobserved for all three treatments (Treatments B, C, and D) vs. TreatmentA (FIG. 1 and Table 2-4).

TABLE 2-4 Effect of Food on Plasma PK Parameters of Compound 1 (N = 31)Treat- ment Ratio of Treat- Geometric Compar- Geometric 90% CI ofParameter ment LS Mean ison LS Mean (%) Ratio (%) AUC₀₋₂₄, A 7.4 mcg*h/LB 6.2 B/A 83.4 74.9, 92.7 C 4.3 C/A 57.7 51.9, 64.2 D 2.8 D/A 37.3 33.6,41.5 AUC_(0-t), A 7.4 mcg*h/L B 6.2 B/A 83.3 74.9, 92.7 C 4.3 C/A 57.751.9, 64.1 D 2.8 D/A 37.9 33.5, 41.4 AUC_(0-inf), A* 10.6 mcg*h/L B 9.0B/A 84.7 75.8, 94.6 C 6.2 C/A 58.4 52.3, 65.3 D 4.0 D/A 37.9 34.0, 42.3C_(max), A 0.66 mcg/L B 0.56 B/A 84.5 75.9, 94.1 C 0.39 C/A 60.1 54.0,66.9 D 0.27 D/A 40.7 36.5, 45.2 *N = 30, a terminal mono-exponentialphase could not be identified for one subject. CI, confidence interval;LS, least squares

The effect of food was more pronounced when a high-fat meal was consumedcloser to dosing and when dairy was included in the meal. Compared witha fasted dose, Compound 1 exposure (C_(max) and AUC) was Reduced by 15%to 17% for a nondairy meal 4 hours before dosing; reduced by 40% to 42%for a nondairy meal 2 hours before dosing; and reduced by 59% to 63% fora dairy meal 2 hours before dosing. The between-subject variability insystemic exposure to omadacycline was similar for Treatments A, B and C(CV 22.4-29.2%) for C_(max) and AUC. By contrast, for Treatment D the CVwas 42.6-44.4% for these parameters.

Concerning safety and tolerability, two subjects experienced treatmentemergent AEs (one reported nausea, one reported somnolence); both eventswere of mild intensity and considered unrelated to study drug. Nosubject discontinued the study for an AE, and no subject experienced aserious AE (SAE). A slight increase from baseline in heart rate (median8 to 10 bpm at 4 to 6 hours post dose) was observed for Treatment A(i.e., the group with highest omadacycline exposure). In all othertreatment groups, the median change from baseline in heart rate was ≤3bpm at all measured time points. No notable changes in blood pressurewere observed. There were no clinically significant changes in clinicallaboratory tests.

The results showed that a single oral dose of Compound 1 was welltolerated. Administration of a 300 mg dose within 2 to 4 hours of foodreduced the bioavailability compared with the fasted state. Thuspreferably, once daily oral Compound 1 should be administered at least 6hours following a meal.

Example 3 A Phase 3 Randomized, Double-Blind, Multi-Center Study toCompare the Safety and Efficacy of Compound 1 IV/PO to MoxifloxacinIV/PO for Treating Adult Subjects with Community-Acquired BacterialPneumonia (CABP)

This study evaluates the safety and efficacy of intravenous (iv) andoral (po) Compound 1 as compared to iv and po moxifloxacin in thetreatment of adults with CABP.

More specifically, the primary objective of this study is to demonstratethat Compound 1 100 mg iv every 12 hours (q12h) for 2 doses, followed by100 mg iv/300 mg po once every 24 hours (q24h) is non-inferior tomoxifloxacin 400 mg iv/po q24h in the treatment of adults with CABP. Thesecondary objectives are: to evaluate the safety of Compound 1 in thetreatment of adult subjects with CABP in the Safety population; toevaluate the Clinical Response according to the identified causativepathogen; and to evaluate the pharmacokinetics (PK) of Compound 1 inadult subjects with CABP.

According to the study design, a randomized (1:1), activecomparator-controlled, double-blind, Phase 3 study was conducted tocompare Compound 1 and moxifloxacin in the treatment of adults with CABP(Pneumonia Outcomes Research Team [PORT] Risk Class II, III, or IV).PORT Risk Class Calculation was adapted from Fine et al., N. Engl. J.Med. 336:243-250, 1997 (incorporated by reference). About 750 patientswere enrolled. Both iv and po phases of the study were double-blind.Enrollment of subjects with disease characterized as PORT Risk Class IIwere limited to no more than 15% of randomized subjects. Enrollment ofsubjects who have received a single dose of an allowed short-actingantibiotic within the 72 hours prior to the first dose of test articlewere limited to no more than 25% of randomized subjects. Enrolledsubjects participated in the study for approximately 30 days.

The study consists of 3 phases: Screening, Double-Blind Treatment, andFollow-up. Screening evaluations, with the exception of the bloodculture sample collection and radiographic confirmation of pneumonia,were completed within the 24 hours prior to randomization. The bloodculture sample collection and radiographic confirmation of pneumoniawere completed within the 24 hours prior to the first dose of testarticle. Following Screening, eligible subjects who meet inclusioncriteria, and do not meet exclusion criteria were randomly assigned to atreatment group to receive 7 to 14 days of treatment with eitherCompound 1 or moxifloxacin. The randomized subjects received their firstdose of test article within 4 hours after randomization.

The comparator drug for this study is selected to be moxifloxacin (400mg iv q24h with the option to transition to 400 mg po q24h), given thewide acceptance of fluoroquinolone monotherapy as a safe, first-lineoption for treating subjects with CABP. Moxifloxacin provides a broadspectrum of activity against respiratory pathogens that are causativeagents of CABP, including typical (e.g., Streptococcus pneumoniae) andatypical (e.g., Legionella, Chlamydophila, and Mycoplasma spp.)pathogens, with a similar spectrum of activity to that of Compound 1.Like Compound 1, moxifloxacin has both iv and po formulation options andwas administered once daily.

A post therapy evaluation visit occurred approximately 5 to 10 daysafter the last dose of test article and a follow-up telephone contactoccurred approximately 30 to 37 days after the first dose of testarticle. Details of the study are further described below.

As used herein, “post therapy evaluation,” “post treatment evaluation,”and “PTE,” are used interchangeably herein throughout the applicationwithout distinction in meaning.

Dosing Regimen:

Compound 1 was administered as 100 mg iv q12h for 2 doses, followed by100 mg iv q24h (starting 24 h after first dose), with the option toswitch to 300 mg po q24h after a minimum of 3 days (4 doses) of ivtreatment.

The comparator drug Moxifloxacin was administered as 400 mg iv q24h(with a single placebo infusion to match the Compound 1 dosing regimen12 hours after the first dose on Day 1), with the option to switch to400 mg po q24h after a minimum of 3 days (4 doses) of iv treatment.

Patient Inclusion/Exclusion Criteria

Patients were male or female 18-years or older. Patients had fulfilledALL of the following criteria: had at least 3 of the following symptoms:cough, production of purulent sputum, dyspnea (shortness of breath), andpleuritic chest pain; had at least two of the following abnormal vitalsigns: fever or hypothermia documented by the investigator(temperature >38.0° C. [100.4° F.] or <36.0° C. [95.5° F.]), hypotensionwith systolic blood pressure (SBP)<90 mm Hg, heart rate >90 beats perminute (bpm), and respiratory rate (RR) >20 breaths/minute; had at least1 clinical sign or laboratory finding associated with CABP: hypoxemia(partial pressure of arterial oxygen [PaO₂]<60 mm Hg by arterial bloodgas [ABG] or oxygen saturation <90% by pulse oximetry), physicalexamination findings of pulmonary consolidation (e.g., dullness onpercussion, bronchial breath sounds, or egophony), and an elevated totalwhite blood cell (WBC) count (>12,000 cells/mm³) or leucopenia (WBC<4,000 cells/mm³) or elevated immature neutrophils (>15% band forms,regardless of total peripheral WBC count); hadradiographically-confirmed pneumonia, i.e., new or progressive pulmonaryinfiltrate(s) on chest X-ray (CXR) or chest computed tomography (CT)scan consistent with acute bacterial pneumonia within 24 or 48 hoursprior to the first dose of test article; and had disease categorized asbeing PORT Risk Class II, III, or IV at Screening, and were expected torequire a minimum of at least 3 days of iv therapy for the initialtreatment of CABP.

Females patients had a negative urine pregnancy test at Screening andagreed to comply with using an acceptable method of birth control as perlocal requirements from Screening through post therapy evaluation (PTE).Males agreed to use an acceptable method of birth control with femalepartner(s) and did not donate sperm from Screening through PTE.

Patients with any of the following conditions were not allowed in thetrial: had received one or more dose(s) of a potentially effectivesystemic antibacterial treatment within the 72 hours prior to the firstdose of test article (a subject was considered to have received apotentially effective systemic antibacterial treatment if the pathogenidentified as causing infection was shown to be susceptible to theantibacterial given or, in the circumstance where a pathogen was notidentified, if the antibacterial agent was approved for treatment ofpneumonia or was known to have activity against any of the leadingcauses of CABP (e.g., Streptococcus pneumoniae, Haemophilus influenzae,Moraxella catarrhalis, Staphylococcus aureus, Legionella pneumophila).One exception was that subjects may be eligible despite priorantibacterial therapy if they had been treated with a single dose of ashort-acting antibacterial (i.e., an antibacterial whose standard dosingregimen was more frequent than once per day); was known or suspected tohave CABP caused by a pathogen that may be resistant to either testarticle (e.g., Klebsiella pneumoniae, Pseudomonas aeruginosa,Pneumocystis jiroveci, obligate anaerobes, mycobacteria, fungalpathogens); suspected or confirmed empyema (a parapneumonic pleuraleffusion was not an exclusion criteria) or lung abscess; subjects withknown or suspected hospital-acquired pneumonia (HAP) orhealthcare-associated pneumonia (HCAP). HAP was defined as pneumoniawith onset of clinical signs and symptoms ≥48 hours afterhospitalization in an acute in-subject health care facility. HCAP wasdefined as pneumonia acquired in a long-term care orsubacute/intermediate healthcare facility (e.g., nursing home) or in asubject admitted with pneumonia following a recent hospitalization(discharged within 90 days of current admission and previouslyhospitalized for ≥48 hours); had known or was clinically suspected tohave 1 or more of the following prior to randomization: alanineaminotransferase (ALT) or aspartate aminotransferase (AST) ≥2×UpperLimit of Normal (ULN), total bilirubin >1.5×ULN, or evidence ofend-stage liver disease (e.g., ascites, hepatic encephalopathy); had aknown history of having experienced unstable cardiac disease (e.g.,unstable angina, myocardial infarction, acute congestive heart failure,unstable cardiac arrhythmia, etc.) within the 3 months prior toScreening; had a QT interval corrected for heart rate using Fridericia'sformula (QTcF) >450 msec (males) or >470 msec (females), were known tohave long QT syndrome, used drugs of potential proarrhythmic or QTprolonging effect, and/or presented with tachyarrhythmia; required anyform of dialysis (e.g., hemodialysis, peritoneal dialysis); history orevidence of severe renal disease or had a calculated creatinineclearance (CrCl) of <30 mL/minute, using the Cockcroft-Gault equation;evidence of significant immunological disease determined by any of thefollowing: current or anticipated neutropenia defined as <500neutrophils/mm³, known infection with Human Immunodeficiency Virus (HIV)and a cluster of differentiation 4 (CD4) count that was unknown ordocumented to be <200 cells/mm³ within the last year, or an AcquiredImmune Deficiency Syndrome (AIDS)-defining illness; the receipt ofcancer chemotherapy, radiotherapy, or potent, non-corticosteroidimmunosuppressant drugs (e.g., cyclosporine, azathioprine, tacrolimus,immune-modulating monoclonal antibody therapy, etc.) within the past 3months, or the receipt of corticosteroids equivalent to or greater than40 mg of prednisone per day or for more than 14 days in the prior 30days; required acute pharmacologic intervention to stabilize bloodpressure (BP) and/or adequate tissue perfusion, OR had evidence ofseptic shock defined by ALL of the following: fever or hypothermiadocumented by the investigator (temperature >38.0° C. [100.4° F.] or<36.0° C. [95.5° F.]), heart rate >90 beats/minute, RR>20breaths/minute, WBC>12,000 cells/mm³ or <4,000 cells/mm³ or >10%immature (band) forms, regardless of the total peripheral WBC count,hypotension with SBP<90 mm Hg despite an iv fluid challenge of 20-30cc/kg over a 30 minute period, and perfusion abnormalities that mayincluded, but were not limited to, lactic acidosis (blood lactateconcentration ≥4 mmol/L), oliguria, or acute alteration in mentalstatus; known or suspected primary or metastatic neoplastic lungdisease, aspiration pneumonia, active tuberculosis, cystic fibrosis,bronchiectasis, bronchial obstruction (e.g., post-obstructivepneumonia), chronic neurological disorder preventing clearance ofpulmonary secretions, or severe chronic obstructive pulmonary disease(COPD); pregnant or nursing (breastfeeding) women; had a history ofhypersensitivity or allergic reaction (e.g., anaphylaxis, urticaria,other significant reaction) to any tetracycline (e.g., minocycline,doxycycline or tigecycline) or to any fluoroquinolone antibiotic; had ahistory of pseudotumor cerebri, or prior (within 2 weeks prior toScreening) or planned concomitant use of isotretinoin; had a history ofsystemic lupus erythematosus or lupus-like syndrome; had currentevidence of pancreatitis; had a history of a central nervous systemdisorder that may predispose to seizures or lower the seizure threshold;use of other investigational drugs within 5 half-lives or 30 days priorto Screening, whichever was longer; had previously been treated withCompound 1 or previously enrolled in this study; any planned medicalintervention that might interfere with the ability to comply with thestudy requirements; and had a life expectancy of less than or equal to 3months or any concomitant condition that, in the opinion of theinvestigator, was likely to interfere with evaluation of the response ofthe infection under study, determination of adverse events (AEs), orcompletion of the expected course of treatment.

In addition, no systemic prior or concomitant antibacterial therapy wasallowed, other than a single dose of a short-acting antibacterial,within the 72 hours prior to the first dose of test article. All othermedications not prohibited by the protocol and considered necessary forthe subject's welfare may be administered and/or continued under thesupervision of the investigator.

Dosing Regimens

The double-blind treatment period was up to 14 days in duration.Subjects who met inclusion criteria and did not meet exclusion criteriawere randomly assigned to a treatment group, and received their firstdose of test article within 4 hours after randomization.

The following assessments were done: vital signs, physical examinations(worsening of observations since the Screening examination was recordedas AEs), AEs and SAEs, concomitant treatments, CABP symptom severityscale, microbiological assessments, 12-lead ECG (performed just prior[within 30 minutes] and 30-90 minutes after the start of the firstinfusion of the first and third doses of test article, at the Day 7visit, at the EOT visit, and as otherwise clinically indicated), bloodfor Central Laboratory assessments: hematology, chemistry, pregnancy(for women only), test article administration and accountability,assessment for po switch or need to continue therapy, and investigator'sassessment of clinical response.

Subjects were randomized (1:1) to 1 of the following 2 treatment arms:

a. Investigational therapy: Compound 1 (supplied with tosylate acidcounter ion, sucrose, hydrochloric acid and sodium hydroxide to adjustthe pH), 100 mg iv q12 h (first 2 doses), followed by 100 mg iv q24h(starting 24 hours after first dose), with the option to switch to 300mg (two 150 mg Compound 1 tablets and 1 over-encapsulated placebo tabletmatching moxifloxacin) po q24h after at least 3 days (4 doses) of ivtreatment.

b. Reference therapy: moxifloxacin, 400 mg iv q24h (with a singleplacebo infusion to match the Compound 1 dosing regimen 12 hours afterthe first dose on Day 1) with the option to switch to 400 mg (one 400 mgmoxifloxacin over-encapsulated tablet and 2 placebo tablets matchingCompound 1 tablets) po q24h after at least 3 days (4 doses) of ivtreatment.

The iv treatment phase (minimum of 3 days, 4 doses) followed adouble-dummy design with placebo infusions matched to active Compound 1and moxifloxacin infusions as shown in Table 3-1 below. Infusions ofCompound 1 and matched placebo were administered continuously overapproximately 30 minutes. During the first 24 hours of iv treatment,subjects on the moxifloxacin treatment arm received a placebo infusionto match the t=12 h infusion in the Compound 1 arm as shown in thetable.

Infusions of moxifloxacin and matched placebo were administeredcontinuously over approximately 60 minutes. All infusion start and stoptimes were recorded.

Table 3-1 Treatment Regimens for IV Test Article Infusion Regimen^(a)Compound 1 Arm^(b,c) Moxifloxacin Arm^(b,c) t = 0 h^(d) Compound 1 100mg in 100 mL NS 100 mL NS placebo 250 mL NS placebo moxifloxacin 400 mgin 250 mL 0.8% saline t= 12 h Compound 1 100 mg in 100 mL NS 100 mL NSplacebo t = 24 h^(d) Compound 1 100 mg in 100 mL NS 100 mL NS placebo250 mL NS placebo moxifloxacin 400 mg in 250 mL 0.8% saline t = 48 h^(d)Compound 1 100 mg in 100 mL NS 100 mL NS placebo 250 mL NS placebomoxifloxacin 400 mg in 250 mL 0.8% saline t = 72 h^(e), Compound 1 100mg in 100 mL NS 100 mL NS placebo then q24 h^(d) 250 mL NS placebomoxifloxacin 400 mg in 250 mL 0.8% saline t = time; NS = Normal saline(0.9% sodium chloride) for injection; q12 h = every 12 hours; q24 h =every 24 hours. ^(a)The start time of the first infusion was designatedtime 0 (t = 0 h), followed by 2 q12 h doses (t = 12 h, t = 24 h), andthen all subsequent doses were q24 h for a minimum of 3 days, 4 doses ofiv treatment (through t = 48 h). ^(b)All 100 mL infusions of Compound 1or 100 mL NS placebo were administered continuously over 30 minutes (atleast 30 minutes and not more than 45 minutes). ^(c)All 250 mL infusionsof moxifloxacin or 250 mL NS placebo were administered continuously overapproximately 60 minutes. ^(d)At these time points a 100 mL infusion wasadministered first, followed by a 250 mL infusion. ^(e)Beginning withthe fifth dose (t = 72 h), based on the investigator decision thetherapy could be iv or was switched to po therapy. Note, the first podose should be administered in the morning, 12-24 hours after the lastiv dose, therefore the first po dose might have occurred as early as t =60 h.

A representative (but none limiting) illustration of the above treatmentregimen for Compound 1 is provided below:

Day 1 Day 2 Day 3 Day 4 and Beyond 0 hr 12 hr 24 hr 48 hr 72 hr - Com-100 100 100 100 After Day 3, either pound 1 mg IV mg IV mg IV mg IV 100mg IV or 300 mg oral q24 h Total duration of treatment (IV + oral (ifpresent)) is typically 7-14 days.

While the subject was receiving iv therapy, the investigator assessedthe subject daily and chose ONE of the following based on the overallclinical assessment of the subject:

(1) continue iv test article;

(2) switch to po test article (after a minimum of 3 days [4 doses] of ivtherapy) Note, the first po dose was administered in the morning, 12-24h after the last iv dose, therefore the first po dose may occur as earlyas t=60 h;

(3) discontinue test article—this decision prompted the EOT evaluation.

Each daily decision was recorded.

At all times during the study the decision to continue iv, switch to po,or discontinue test article was made based on the clinical judgment ofthe investigator. The investigator may have used the culture andsusceptibility results from the local microbiology laboratory to helpguide therapy; however, decisions to continue or discontinue testarticle was based on clinical response rather than susceptibilityresults (as Compound 1 susceptibility testing was not available at thelocal site). If the CABP was caused by a microorganism that was notsusceptible to moxifloxacin in vitro, the decision to continue ordiscontinue study treatment was based on the subject's clinical courseand the investigator's clinical judgment. The rationale for thisdecision was recorded.

The decision to switch to po treatment was made by the investigator. Fora subject to be considered clinically stable and meet criteria fortransition to a po regimen, they must have had the following findingsnoted and recorded:

a. Temperature ≤37.8° C. (100° F.)

b. Heart rate ≤100 beats/minute

c. RR≤24 breaths/minute

d. SBP ≥95 mm Hg

e. Oxygen saturation ≥90% as measured by pulse oximetry or PaO₂≥60 mm Hgby ABG

f. No worsening of CABP symptoms (cough, sputum production, pleuriticchest pain, dyspnea) compared to Screening

g. Normal mental status (“absence of confusion” or pre-illness Baselinefor subjects who did not have normal mental status before onset ofpneumonia)

h. Ability to maintain po intake.

Switch to po was not permitted until after the subject had completed atleast the first 3 days of iv treatment (after 4 iv doses).

The date and time the investigator confirmed the criteria for thesubject's eligibility for po treatment were met and made the decision toswitch to po treatment was recorded. For subjects that had been switchedto po test article and discharged from the hospital prior to study Day6, visits were conducted on study Days 4 and 5, while a study Day 6visit was optional.

Treatment regimens for po dosing were shown in Table 3-2. When switchingfrom iv to po test article the recommended interval between doses wasmaintained. The first po dose, for both Compound 1 and moxifloxacintreatment arms, was given in the morning 12 to 24 hours after the lastiv dose. The po treatment phase also employed a double-blind,double-dummy design using Compound 1 placebo comparator tablets ofmatching size and shape to active Compound 1 tablets and matchingover-encapsulated placebo and active moxifloxacin tablets.

The tablets had excipients including lactose monohydrate,microcrystalline cellulose, sodium stearyl fumarate, crospovidone,colloidal silicone dioxide, sodium bisulfite, polyvinyl alcohol,titanium dioxide, talc, soya lecithin, xanthan gum, FD&C Yellow #6 andFD&C #2. The Compound 1 tablets were taken with water in a fasting state(no food, antacids or multivitamins containing multivalent cations(e.g., aluminum, magnesium, calcium, bismuth, iron, or zinc) or drinkexcept water for at least 6 hours). After dosing, no food was to beconsumed for 2 hours; no dairy products, antacids or multivitaminscontaining multivalent cations (e.g., aluminum, magnesium, calcium,bismuth, iron, or zinc) for 4 hours.

TABLE 3-2 Treatment Regimens for Oral Test Article a) Time b) Compound 1c) Moxifloxacin d) Dosing of Dosing Arm Arm Condition^(a, b) Morning Two150 mg One 400 mg Fasting overnight (no tablets and 1 over-encapsulatedfood or drink except over-encapsulated tablet and 2 water for at least 6placebo tablet placebo tablets hours) before dosing; resemblingresembling after dosing, no food moxifloxacin Compound 1 for 2 hours, nodairy tablets products for 4 hours ^(a)All doses of po test article weretaken with water. ^(b) All subjects were instructed to avoid takingantacids and multivitamins containing multivalent cations (e.g.,aluminum, magnesium, calcium, bismuth, iron, or zinc) while taking potest article.

While the subject was receiving po therapy, the investigator assessedthe subject on study Day 7, 10 and 14 and choose 1 of the followingactions:

a. continue po test article;

b. discontinue test article—this decision prompted the EOT evaluation.

The investigator may have used the culture and susceptibility resultsfrom the local microbiology laboratory to help guide therapy; however,decisions to continue or discontinue test article was based on clinicalresponse rather than susceptibility results (as Compound 1susceptibility testing was not available at the local site). If the CABPwas caused by a microorganism that was not susceptible to moxifloxacinin vitro, the decision to continue or discontinue study treatment wasbased on the subject's clinical course and the investigator's clinicaljudgment.

No dose adjustments and interruptions of test article were permitted.

Subjects were evaluated at 2 visits after the completion of treatment:at the PTE 5 to 10 days after the last treatment day, and at a FinalFollow-up assessment 30 to 37 days after the first dose of treatment.

The following table summarizes the study drug exposure in the safetypopulation.

Compound 1 Moxifloxacin All Subjects Characteristics (N = 382) (N = 388)(N = 770) Duration of exposure on therapy (days) n 382 388 770 Mean (SD)9.6 (2.88) 9.6 (2.94) 9.6 (2.91) Medium 10.0 10.0 10.0 Min, Max 1, 14 1,17 1, 17 Duration of exposure on therapy (days), n (%) 382 388 770 0 0 00 1-3 17 (4.5) 17 (4.4) 34 (4.4) 4-6 10 (2.6) 17 (4.4) 27 (3.5) 7-10 238(62.3) 218 (56.2) 456 (59.2) 11-14 117 (30.6) 134 (34.5) 251 (32.6) >140 2 (0.5) 2 (0.3) Number of days of IV therapy n 382 388 770 Mean (SD)5.7 (2.51) 5.7 (2.54) 5.7 (2.52) Medium 5.0 5.0 5.0 Min, Max 1, 14 1, 141, 14 Number of days of IV therapy, n (%) 382 388 770 0 0 0 0 1-2 15(3.9) 13 (3.4) 28 (3.6) 3-6 222 (58.1) 232 (59.8) 454 (59.0) 7-10 134(35.1) 130 (33.5) 264 (34.3) 11-14 11 (2.9) 13 (3.4) 24 (3.1) >14 0 0 0Number of days of IV therapy prior to oral switch n 295 294 589 Mean(SD) 5.0 (2.02) 5.0 (2.08) 5.0 (2.04) Medium 4.7 4.7 4.7 Min, Max 2, 132, 11 2, 13 Subjects switching to oral therapy, n (%) 295 (77.2) 294(75.8) 589 (76.5) Day of oral switch, n (%) 295 294 589 1-3 1 (0.3) 1(0.3) 2 (0.3) 4-5 130 (44.1) 132 (44.9) 262 (44.5) 6-7 80 (27.1) 81(27.6) 161 (27.3) ≥8 84 (28.5) 80 (27.2) 164 (27.8) Number of days oforal therapy n 295 294 589 Mean (SD) 5.1 (1.93) 5.2 (2.04) 5.2 (1.98)Medium 5.0 5.0 5.0 Min, Max 1, 11 1, 14 1, 14 Number of days of oraltherapy, n (%) 295 294 589 0 0 0 0 1-4 132 (44.7) 132 (44.9) 264 (44.8)5-7 152 (51.5) 147 (50.0) 299 (50.8) 8-11 11 (3.7) 14 (4.8) 25 (4.2) >110 1 (0.3) 1 (0.2) Note: Both active test article and placebo areincluded.

Percentages for IV summaries are based on subjects receiving at leastone IV dose. Percentages for oral summaries are based on subjectsreceiving at least one oral dose. Percentages for subjects who switchedfrom IV to oral are based on the number who switched (e.g., 295 forCompound 1).

Note: Duration of study drug exposure in days=Date of last IV or oraldose−Date of first IV dose+1. Number of days of IV therapy (IV dosing)prior to oral switch=Date of last IV dose−Date of first IV dose+1.

Criteria for switching from IV to oral treatment include: Temperature≤37.8° C. (100° F.); Heart rate ≤100 beats/minute; Respiratory Rate <=24breaths/minute; Systolic Blood Pressure ≥95 mmHg; Oxygen saturation ≥90%by pulse oximetry or PaO₂≥60 mmHg by ABG; No worsening of CABP symptomscompared to screening; Normal Mental Status; and Ability to maintain POintake. For patients administered Compound 1, 100% of the 295 patientswho switched from IV dose to oral dose satisfied each of the abovecriteria.

Safety Evaluation

Any subject who received test article were included in the evaluationfor safety. Safety evaluations of the enrolled subjects includedphysical exams, vital signs (blood pressure, pulse rate, bodytemperature), AEs and SAEs, laboratory assessments (hematology, serumchemistry, urinalysis), 12-lead electrocardiogram (ECG) evaluations, andpregnancy assessments.

After Screening, a physical examination was conducted on the study daysand at the EOT and PTE visits. Any new clinically significant findingoccurs (i.e., not noted at Screening) after the Screening exam wascaptured as an AE.

Vital signs including body temperature, BP, pulse/heart rate, and RRwere recorded prior to each dose while the subject was on iv treatment.

Blood samples for hematology, chemistry and coagulation (prothrombintime only) were drawn at Screening, Day 4, Day 7, Day 10, EOT, and PTE.

Safety studies included a standard 12-lead ECG at the following times:screening, just prior (within 30 minutes) to the start of the firstinfusion of the first dose of test article (t=0 h), 30-90 minutes afterthe start of the first infusion of the first dose of test article, justprior (within 30 minutes) to the start of the first infusion of thethird dose of test article (t=24 h), 30-90 minutes after the start ofthe first infusion of the third dose of test article, at the Day 7visit, at the EOT visit, and in any case in which a subject developed anAE of non-pleuritic cardiac chest pain, palpitations, tachyarrhythmia oras otherwise clinically indicated.

Safety studies also included pregnancy and assessments of fertility. Allwomen had a urine pregnancy test at the site at the Screening visit. Noone was enrolled if a positive urine pregnancy test result was obtained.A serum sample for β-hCG testing was also collected at the Screeningvisit and sent to the Central Laboratory for confirmation of the urinepregnancy results. Serum samples for β-hCG testing at the CentralLaboratory were also collected at EOT and PTE. Test articleadministration was discontinued if a positive β-hCG result was reportedby the Central Laboratory after a woman was enrolled.

Safety studies also included two sets of blood cultures collected withinthe 24 hours prior to the first dose of test article. Each set of bloodcultures were collected by direct venipuncture from independent bodysites 15-30 minutes apart. If bacteria were isolated from baseline bloodcultures, repeat blood cultures were collected on the day that thepositive blood culture is detected. If subsequent blood cultures werealso positive, the blood cultures were repeated as necessary untilnegative blood cultures were obtained.

Efficacy Evaluation

In order to satisfy different health authority requirements, the primaryvariables were tested with 2 response endpoints:

-   -   Successful Early Clinical Response or ECR (72-120 hours after        first dose) was determined programmatically and defined as        survival with improvement in at least 2 of 4 subject symptoms        (cough, sputum production, pleuritic chest pain, dyspnea), as        assessed by the investigator, without deterioration in any of        these 4 symptoms (FDA primary efficacy point). One such        evaluation was done in the ITT population.    -   Successful Investigator's Assessment of Clinical Response at the        PTE visit, defined as survival after completion of a test        article regimen, with resolution of signs and symptoms of the        infection to the extent that further antibacterial therapy was        not necessary (EMA primary efficacy point). One of such        evaluation was done in both the ITT and CE populations, limited        to PORT Risk Class III/IV subjects. In addition, 97.5% CI was        used in addition to the 95% CI level for 10% non-inferiority        assessment.

The Early Clinical Response endpoint was tested in the intent-to-treat(ITT) analysis population. The Investigator's Assessment of ClinicalResponse at PTE endpoint was tested in the ITT and clinically evaluable(CE) populations (co-primary endpoints).

Secondary efficacy variables included:

-   -   Response category for Early Clinical Response    -   Clinical Response category for Investigator's Assessment of        Clinical Response at EOT and PTE    -   Clinical Response category according to the identified causative        pathogen

The following is a list of key assessments that was performed:

-   -   Assessment of signs and symptoms of CABP by the investigator    -   Microbiological assessment of the infection    -   Assessment of clinical response

Each of the key assessments is described in further detail below.

a) Assessment of CABP Symptom Severity

The assessment of CABP symptoms observed by the investigator wasconducted at every scheduled evaluation with the exception of the FinalFollow-up assessment. The investigator specifically assessed theseverity level of the subject's symptoms of cough, sputum production,pleuritic chest pain and dyspnea on a 4 point scale (absent, mild,moderate, or severe) based upon the Community Acquired BacterialPneumonia Subject Symptom Severity Guidance Framework for InvestigatorAssessment (see below) and entered the symptom severity scores into theeCRF. For subjects that had been switched to po test article anddischarged from the hospital prior to study Day 6, visits were conductedon study Days 4 and 5, while a study Day 6 visit was optional.

CABP Subject Symptom Severity Guidance Framework for InvestigatorAssessment

Absent Mild Moderate Severe COUGH? No cough or Cough present but Coughpresent, Cough is present resolution (to pre- it does not interferefrequent and it does throughout the day CABP Baseline) with subject'susual interfere with some and night; it limits daily activities of thesubject's most of the usual daily subjects' usual activities dailyactivities and sleep patterns PLEURITIC No chest pain or Chest painpresent Chest pain is Chest pain is CHEST PAIN? resolution of chestoccasionally with present with normal present at rest pain related todeep breathing but breaths and it does and/or with shallow CABP it doesnot interfere interfere with the breathing; it limits with subject'susual subject's usual most of the daily activities daily activitiessubject's usual daily activities SHORTNESS OF No shortness of Shortnessof breath Shortness of breath Shortness of breath BREATH? breath orresolution with strenuous with usual activities with minimal (topre-CABP activities only but it and it does interfere exertion or atrest; Baseline) does not interfere with the subject's it limits most ofthe with subject's usual usual daily subject's usual daily activitiesactivities daily activities PHLEGM/ No coughing up of Subject coughs upSubject coughs up Subject coughs up SPUTUM phlegm/sputum or a smallamount of a moderate amount a large amount of PRODUCTION? resolution (topre- phlegm/sputum of phlegm/sputum phlegm/sputum CABP Baseline)

b) Microbiological Assessments

Microbiological assessments included respiratory culture and Gram stain,urine test for Legionella pneumophila and Streptococcus pneumoniaeantigen screening, and serology test for Legionella pneumophila,Mycoplasma pneumoniae and Chlamydophila pneumoniae titers.

For respiratory culture and Gram stain, at the Screening visitcollection of an adequate quality expectorated or induced sputum orother respiratory specimen reflecting fluid from the lower respiratorytract (e.g., respiratory fluid obtained by bronchoalveolar lavage orbronchoscopy; pleural fluid obtained by thoracentesis; or expectoratedor induced sputum meeting adequacy criteria) was attempted from allsubjects and submitted to the local microbiology laboratory for Gramstain and culture. The date, time and type of specimen submitted wererecorded. An adequate quality sputum specimen was defined as having thefollowing 2 findings as reported by the local laboratory:

1. <10 Squamous epithelial cells/low power field (lpf) (i.e., 100×)

2. >25 Polymorphonuclear cells/lpf (i.e., 100×)

Adequate quality sputum specimens and other Screening respiratoryspecimens for culture were obtained prior to first dose of test article.At the EOT and/or PTE visit, respiratory specimen cultures and Gramstains were obtained only for subjects who were Clinical Failures andrequired alternative antibacterial treatment for CABP.

Laboratory reports on Gram stains included a semi-quantitativedescription of the number of polymorphonuclear leukocytes per low powerfield (i.e., 100×) and a description of bacteria seen. For Gram stainsof respiratory specimens a semi-quantitative description of the numberof squamous epithelial cells per low power field (i.e., 100×) wasincluded.

Culture results included identification of all pathogens to the level ofgenus and species. Susceptibility testing for moxifloxacin (or otherfluoroquinolones) was performed using a standard method.

All isolates identified from expectorated or induced sputum specimensmeeting the 2 criteria that define the specimen as being of adequatequality and/or that were isolated from respiratory specimens or bloodand were potential pathogens were submitted to the Central Laboratoryfor verification of genus and species and for standardized minimuminhibitory concentration (MIC) testing performed for Compound 1,moxifloxacin and a panel of currently approved antibiotics.

Regarding urine tests for Legionella pneumophila and Streptococcuspneumoniae antigen screening, urine was collected at the Screening visitto test for the presence of Legionella pneumophila and Streptococcuspneumoniae antigens.

Regarding serology test for Legionella pneumophila, Mycoplasmapneumoniae and Chlamydophila pneumoniae titers, blood samples werecollected to conduct serology for Legionella pneumophila, Mycoplasmapneumoniae and Chlamydophila pneumoniae by the Central Laboratory at theScreening visit, and at the PTE visit.

c) Assessment of Clinical Outcome

Assessment of clinical outcome occurred at Early Clinical Responseassessment (programmatically), EOT, and PTE as described below.

1. Evaluation of the Infection Under Study at the Early ClinicalResponse Assessment

The formal determination of the response to therapy at the EarlyClinical Response assessment (72 to 120 hours after administration ofthe first dose of test article) was done programmatically using theinvestigator's assessment of the subject's symptoms associated with CABPentered into the eCRF. The investigator was not responsible forcategorizing subjects as Clinical Success, Failure, or Indeterminate atthe Early Clinical Response assessment. The severity of the subject CABPsymptoms of cough, sputum production, pleuritic chest pain and dyspneawas evaluated on a 4-point scale (absent, mild, moderate, or severe)based upon the Community Acquired Bacterial Pneumonia Subject SymptomSeverity Guidance Framework for Investigator Assessment. A CABP subjectsymptom severity assessment was completed at every scheduled evaluationwith the exception of the Final Follow-up assessment. For subjects thathad been switched to po test article and discharged from the hospitalprior to study Day 6, visits was conducted on study Days 4 and 5, whilea study Day 6 visit was optional.

Clinical Success: at the Early Clinical Response assessment was definedas survival with improvement of at least 1 level (i.e., severe tomoderate, moderate to mild, mild to absent) compared to Screening in 2CABP symptoms (cough, sputum production, pleuritic chest pain anddyspnea) with no worsening by at least 1 level in the other inclusionCABP symptoms. In order for the subject to be considered a ClinicalSuccess, the subject may not meet any criteria for Clinical Failure orIndeterminate Early Clinical Response.

Clinical Failure: defined as meeting any of the following criteria:

-   -   There was no improvement by at least 1 level (i.e., severe to        moderate, moderate to mild, mild to absent) compared to        Screening in 2 CABP symptoms.    -   Any of the 4 CABP symptoms was worse (by at least 1 level)        compared to Screening.    -   The subject required alternative (rescue) antibacterial        treatment for CABP prior to the Early Clinical Response        assessment related to either (a) progression or development of        new symptoms attributable to CABP or (b) development of        infectious complications of CABP (e.g., empyema, lung abscess).    -   The subject was receiving antibacterial therapy that may be        effective for the infection under study for a different        infection from the 1 under study.    -   Discontinued study therapy due to an AE and received alternative        antibacterial treatment for CABP prior to the Early Clinical        Response assessment.    -   Death prior to the Early Clinical Response assessment.

Indeterminate: the clinical response to test article can not beadequately inferred due to:

-   -   Subject was not seen for the evaluation because they withdrew        consent, were lost to follow-up, other reason (specify).    -   Other specified reason.

2. Clinical Evaluation of the Infection Under Study at EOT

EOT assessments were performed on the calendar day of, or within 2 daysfollowing the last dose of any test article. If a subject withdrewprematurely or terminated participation in the study prior to completionof the planned antibiotic therapy, the EOT visit was conducted.

The investigator determined whether or not the subject met the criteriaof 1 of the following clinical outcomes:

Clinical Success: the subject was alive and the infection wassufficiently resolved such that further antibacterial therapy was notneeded. These subjects may have some residual findings related toinfection (i.e., cough) requiring ancillary (i.e., non-antibiotic)treatment (e.g., expectorant). In order for the subject to be considereda Clinical Success at EOT, the subject may not meet any criteria forClinical Failure or Indeterminate at EOT.

Clinical Failure: the subject required alternative antibacterialtreatment for CABP prior to EOT related to either (a) progression ordevelopment of new symptoms of CABP or (b) development of infectiouscomplications of CABP (e.g., empyema, lung abscess) or (c) subjectdeveloped an AE that required discontinuation of study therapy. Otherreasons for Clinical Failure are:

-   -   Subject was receiving antibacterial therapy that may be        effective for the infection under study for a different        infection from the 1 under study.    -   Death prior to EOT visit.

Indeterminate: the clinical response to test article could not beadequately inferred due to:

-   -   Subjects were not seen for EOT evaluation because they withdrew        consent, were lost to follow-up, other reason (specify).    -   Other specified reason.

3. Clinical Evaluation of the Infection Under Study at PTE

The PTE assessment was performed 5 to 10 days after the subject's lastday of therapy. The investigator determined whether or not the subjectmet the criteria of 1 of the following clinical outcomes:

Clinical Success: survival after completion of a test article regimenwithout receiving any systemic antibacterial therapy other than testarticle, resolution of signs and symptoms of the infection present atScreening with no new symptoms or complications attributable to CABP andno need for further antibacterial therapy.

Clinical Failure: the subject required alternative antibacterialtreatment for CABP prior to PTE related to either (a) progression ordevelopment of new symptoms of CABP or (b) development of infectiouscomplications of CABP (e.g., empyema, lung abscess).

-   -   The subject was receiving antibiotics that may be effective for        the infection under study for a different infection from the 1        under study.    -   Death prior to PTE.

Indeterminate: the clinical response to test article could not beadequately inferred due to:

-   -   Subjects were not seen for PTE evaluation because they withdrew        consent, were lost to follow-up, other (specify).    -   Other specified reason.

d) Pharmacokinetic Plasma Samples for Compound 1 Concentration

PK data was analyzed using a population PK model. PK samples werecollected using a sparse sampling method for the population PK model.The number of samples and collection schedule varied for individualsubjects. Up to 4 samples were collected per subject between study Days1 to 7. Blood was collected either by fresh venipuncture or via acannula used SOLEY for that purpose (PK samples were NOT drawn throughthe same iv access used for administration of test article). The datesand times for all doses of test article and PK sample collections wererecorded. For intravenously administered doses of test article, thestart and stop times for each infusion were recorded. The sample tubewere centrifuged at 1500×g for 10 minutes and the separated plasmatransferred in 2 equal aliquots into pre-labeled tubes; and the tubesfrozen at −70° C. within 60 minutes of collection. The time the samplewas frozen was recorded to the nearest minute. All of the PK samplesfrom a single subject were collected and frozen at −70° C. and shippedfrozen at −70° C. to the Central Laboratory. The samples were assayed atAnalytical Laboratory for Compound 1 using a specific, sensitive andvalidated Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS)method.

e) Safety Monitoring—Adverse Events

An AE was defined as any untoward, undesired, or unplanned event in theform of signs, symptoms, disease, or laboratory or physiologicobservations occurring in a person given a test article or in a clinicalstudy. The event does not need to be causally related to the testarticle or clinical study. An AE included, but was not limited to, thefollowing:

-   -   Any clinically significant worsening of a preexisting condition.    -   An AE occurring from overdose of a test article, whether        accidental or intentional. Overdose is a dose greater than that        specified in the protocol.    -   An AE occurring from abuse (e.g., use for nonclinical reasons)        of a test article.    -   An AE that had been associated with the discontinuation of the        use of a test article. A SAE is an AE that:    -   Resulted in death.    -   Was life-threatening (see below).    -   Required hospitalization or prolongation of an existing        hospitalization (see below).    -   Resulted in a persistent or significant disability or incapacity        (see below).    -   Resulted in cancer.    -   Resulted in a congenital anomaly or birth defect.    -   Additionally, important medical events that may not result in        death, be life-threatening, or require hospitalization may be        considered SAEs when, based on appropriate medical judgment,        they may jeopardize the subject and may require medical or        surgical intervention to prevent 1 of the outcomes listed in        this definition. Examples of such events included allergic        broncho spasm requiring intensive treatment in an emergency room        or at home, blood dyscrasias or convulsions that did not result        in hospitalization, or development of drug dependency or drug        abuse.

A protocol-related AE was defined as an AE occurring during a clinicalstudy that was not related to the test article, but was considered bythe investigator or the Medical Monitor (or designee) to be related tothe research conditions, i.e., related to the fact that a subject wasparticipating in the study. For example, a protocol-related AE may be anuntoward event related to a medical procedure required by the protocol.

Certain information, although not considered an SAE, must be recorded,reported, and followed up as indicated for an SAE. This included:pregnancy exposure to a test article, lactation exposure to a testarticle with or without an AE, overdose of a test article as specifiedin this protocol with or without an AE, and inadvertent or accidentalexposure to a test article with or without an AE.

Data Analysis

All analyses of data for this study complied with InternationalConference on Harmonisation of Technical Requirements for Registrationof Pharmaceuticals for Human Use (ICH-E9) and the sponsor's guidancedocuments and standards. Statistical analyses were performed usingStatistical Analysis Software (SAS).

a) Analysis Populations

A number of subject populations had been defined for the variousanalyses of efficacy and safety, as follows:

-   -   The ITT population consisted of all randomized subjects.    -   The microbiological intent-to-treat (microlTT) population        consisted of subjects in the ITT population who had at least 1        causative pathogen identified at Screening from culture of a        respiratory specimen (e.g., respiratory fluid obtained by        bronchoalveolar lavage or bronchoscopy; pleural fluid obtained        by thoracentesis; or expectorated or induced sputum meeting        adequacy criteria), culture of blood, or from a        culture-independent method (e.g., positive urinary antigen test        for Streptococcus pneumoniae or Legionella pneumophila, or        positive serology for Legionella pneumophila, Mycoplasma        pneumoniae or Chlamydophila pneumoniae).    -   Expanded microlTT population was defined using the same criteria        as for the microlTT population, except that an adequate Gram        stain was defined as >10 PMNs/LPF and <10 SECs/LPF for        determination of whether an isolate from a sputum culture is a        pathogen or not. Here, PMN=Polymorphonuclear; SEC=Squamous        Epithelial Cells; LPF=Low Power Field.    -   The CE population consisted of all ITT subjects who received        test article, had a qualifying CABP, an assessment of outcome,        and met all other evaluability criteria detailed in the SAP.    -   The CE-EOT/PTE population consisted of all randomized safety        subjects who received any amount of active test article, had        completed the investigator's assessment of clinical response at        the EOT/PTE visit, with no indeterminate clinical response and        met specific criteria related to the required assessments. Here,        CE=Clinically Evaluable; EOT=End of treatment; PTE=Post Therapy        Evaluation.    -   The microbiologically evaluable (ME) population included        subjects in the CE population who have at least 1 causative        pathogen at Screening.    -   The ME-EOT/PTE consisted of all subjects in both microlTT and        the CE-EOT/PTE populations.    -   The Safety population consists of all randomized subjects who        receive test article.

The various study populations enrolled in the study, as defined above,are listed below.

Compound 1 Moxifloxacin All Subjects Population n (%) n (%) n (%) ITT386 388 774 Safety 382 (99.0)  388 (100.0) 770 (99.5) microITT 204(52.8) 182 (46.9) 386 (49.9) Expanded microITT 218 (56.5) 198 (51.0) 416(53.7) CE-EOT 357 (92.5) 357 (92.0) 714 (92.2) CE-PTE 340 (88.1) 345(88.9) 685 (88.5) ME-EOT 193 (50.0) 172 (44.3) 365 (47.2) ME-PTE 188(48.7) 169 (43.6) 357 (46.1)

b) Subject Demographics and Baseline Characteristics

Descriptive statistics, by treatment arm, was provided for thefollowing: Subject disposition (completed test article, discontinuedtest article by reason for discontinuation, completed study, anddiscontinued study by reason for discontinuation); Protocol deviations;CABP background information (subject demographics: age (years), gender,race, height (cm), weight (kg), Body Mass Index (BMI) (kg/m²)); PORTRisk Class; and Medical histories and continuing medical conditions.

Baseline demographic and medical variables were analyzed using a 2-sidedFisher's exact test (for categorical variables) or a 2-sided WilcoxonRank Sum test (for ordinal and continuous variables).

Selected results of these analysis were compiled in the tables below.

Subject Disposition - ITT Population Compound 1 Moxifloxacin AllSubjects (N = 386) (N = 388) (N = 774) Parameter/Category n (%) n (%) n(%) p-value Randomized 386 (100.0) 388 (100.0) 774 (100.0) CompletedStudy Treatment^([1]) 352 (91.2) 346 (89.2) 698 (90.2) PrematurelyDiscontinued from Study 34 (8.8) 42 (10.8) 76 (9.8) 0.3981 TreatmentReason For Premature Discontinuation from Study Treatment Adverse Event17 (4.4) 28 (7.2) 45 (5.8) Lost to Follow-up 0 1 (0.3) 1 (0.1)Withdrawal by Subject 4 (1.0) 3 (0.8) 7 (0.9) Physician Decision 3 (0.8)9 (2.3) 12 (1.6) Death 4 (1.0) 1 (0.3) 5 (0.6) Other 6 (1.6) 0 6 (0.8)Completed Study^([2]) 356 (92.2) 362 (93.3) 718 (92.8) PrematurelyDiscontinued from Study 30 (7.8) 26 (6.7) 56 (7.2) 0.5819 Reason ForPremature Discontinuation from Study Adverse Event 7 (1.8) 9 (2.3) 16(2.1) Lost to Follow-up 0 3 (0.8) 3 (0.4) Withdrawal by Subject 7 (1.8)8 (2.1) 15 (1.9) Physician Decision 0 1 (0.3) 1 (0.1) Death 6 (1.6) 3(0.8) 9 (1.2) Other 10 (2.6) 2 (0.5) 12 (1.6) ^([1])Summary of subjectsthat completed the study treatment. ^([2])Summary of subjects thatcompleted the study (i.e., received at least one dose of test articleand completed EOT, PTE and Follow-up).

Percentages are based on the ITT population. P-values for differencesbetween treatment groups are from Fisher's exact test. Number ofsubjects prematurely discontinuing study treatment due to an adverseevent may not match counts in adverse events tables as some reasons fordiscontinuations are coded as death rather than adverse event. Subjectsrandomized but not treated (total n=4) are counted in the Othercategory. Total number of deaths are 12 (8 omadacycline and 4moxifloxacin). EOT=End of Treatment; PTE=Post Treatment Evaluation.

Subject Disposition - CE-PTE Population Compound 1 Moxifloxacin AllSubjects (N = 340) (N = 345) (N = 685) Parameter/Category n (%) n (%) n(%) p-value Randomized 340 (100.0) 345 (100.0) 685 (100.0) CompletedStudy Treatment^([1]) 322 (94.7) 317 (91.9) 639 (93.3) PrematurelyDiscontinued from Study 18 (5.3) 28 (8.1) 46 (6.7) 0.1694 TreatmentReason For Premature Discontinuation from Study Treatment Adverse Event12 (3.5) 19 (5.5) 31 (4.5) Lost to Follow-up 0 0 0 Withdrawal by Subject0 0 0 Physician Decision 3 (0.9) 8 (2.3) 11 (1.6) Death 3 (0.9) 1 (0.3)4 (0.6) Other 0 0 0 Completed Study^([2]) 330 (97.1) 339 (98.3) 669(97.7) Prematurely Discontinued from Study 10 (2.9) 6 (1.7) 16 (2.3)0.3234 Reason For Premature Discontinuation from Study Adverse Event 4(1.2) 2 (0.6) 6 (0.9) Lost to Follow-up 0 0 0 Withdrawal by Subject 1(0.3) 0 1 (0.1) Physician Decision 0 1 (0.3) 1 (0.1) Death 5 (1.5) 3(0.9) 8 (1.2) Other 0 0 0 ^([1])Summary of subjects that completed thestudy treatment. ^([2])Summary of subjects that completed the study(i.e., received at least one dose of test article and completed EOT, PTEand Follow-up).

Percentages in the table above are based on the CE-PTE population.p-values for differences between treatment groups were from Fisher'sexact test. The number of subjects prematurely discontinuing studytreatment due to an adverse event may not match counts in adverse eventstables as some reasons for discontinuations were coded as death ratherthan adverse event.

Demographic and Baseline Characteristics - Safety Population Compound 1Moxifloxacin All Subjects Characteristics (N = 382) (N = 388) (N = 770)p-value Gender, n (%) n 382 388 770 Female 177 (46.3) 169 (43.6) 346(44.9) Male 205 (53.7) 219 (56.4) 424 (55.1) 0.4689 Race, n (%) n 382388 770 White 353 (92.4) 355 (91.5) 708 (91.9) Black or African 11 (2.9)7 (1.8) 18 (2.3) American Asian 17 (4.5) 18 (4.6) 35 (4.5) AmericanIndian or 0 2 (0.5) 2 (0.3) Alaska Native Native Hawaiian or 0 0 0 OtherPacific Islander Other 1 (0.3) 6 (1.5) 7 (0.9) 0.1762 Ethnicity, n (%) n382 388 770 Hispanic or Latino 8 (2.1) 14 (3.6) 22 (2.9) Not Hispanic orLatino 370 (96.9) 370 (95.4) 740 (96.1) Not Reported/Unknown 4 (1.0) 4(1.0) 8 (1.0) 0.4465 Age (years) n 382 388 770 Mean (SD) 60.9 (15.18)62.1 (15.21) 61.5 (15.20) Median 61.0 63.0 62.0 Min, Max 19, 97 19, 9419, 97 0.1394 Categorical Age (years), n (%) n 382 388 770 18-45 61(16.0) 61 (15.7) 122 (15.8) >45-65 171 (44.8) 155 (39.9) 326 (42.3) >65150 (39.3) 172 (44.3) 322 (41.8) 0.3303 >75 74 (19.4) 83 (21.4) 157(20.4) Height (cm) n 382 388 770 Mean (SD) 168.6 (9.90) 168.5 (9.84)168.6 (9.87) Median 169.0 168.0 169.0 Min, Max 137, 196 135, 198 135,198 0.8528 Weight (kg) n 382 388 770 Mean (SD) 77.67 (18.018) 78.00(17.861) 77.83 (17.928) Median 76.00 77.50 77.00 Min, Max  36.0, 147.0 28.0, 145.2  28.0, 147.0 0.9687 BMI (kg/m²) n 382 388 770 Mean (SD)27.26 (5.763) 27.42 (5.791) 27.34 (5.774) Median 26.28 26.50 26.36 Min,Max 16.0, 50.6 12.6, 54.7 12.6, 54.7 0.6103 Renal Function (Local Lab),n (%) n 382 388 770 Normal renal function 186 (48.7) 207 (53.4) 393(51.0) [CrCl >80 mL/min] Mild renal impairment 127 (33.2) 119 (30.7) 246(31.9) [CrCl >50-80 mL/min] Mod. renal impairment 69 (18.1) 62 (16.0)131 (17.0) [CrCl 30-50 mL/min] Severe renal impairment 0 0 0 0.4245[CrCl <30 mL/min] PORT Score (actual) n 382 388 770 Mean (SD) 83.1(16.25) 84.0 (15.99) 83.5 (16.12) Median 80.0 82.0 81.0 Min, Max  48,130  36, 136  36, 136 0.1571 PORT Risk Class (actual) n 382 388 770 I (0≤ Port Score ≤ 50) 2 (0.5) 2 (0.5) 4 (0.5) II (51 ≤ PortScore ≤ 70) 54(14.1) 54 (13.9) 108 (14.0) III (71 ≤ Port Score ≤ 90) 226 (59.2) 216(55.7) 442 (57.4) IV (91 ≤ Port Score ≤ 130) 100 (26.2) 115 (29.6) 215(27.9) V (Port Score ≥ 131) 0 1 (0.3) 1 (0.1) 0.7341

In the table above, Age is calculated from the date of birth to theinformed consent date. p-values for differences between treatment groupsare from Fisher's exact test (for categorical variables) or WilcoxonRank Sum test (for continuous variables). For each categoricalparameter, the denominator for the percentage is the number of subjectswho had that parameter assessed. PORT Score (actual) and PORT Risk Class(actual) are based on PORT score (derived/corrected) from CRF.

Demographic and Baseline Characteristics - CE-PTE PopulationOmadacycline Moxifloxacin All Subjects Characteristics (N = 340) (N =345) (N = 685) p-value Gender, n (%) n 340 345 685 Female 154 (45.3) 152(44.1) 306 (44.7) Male 186 (54.7) 193 (55.9) 379 (55.3) 0.7590 Race, n(%) n 340 345 685 White 312 (91.8) 316 (91.6) 628 (91.7) Black orAfrican American 11 (3.2) 6 (1.7) 17 (2.5) Asian 16 (4.7) 17 (4.9) 33(4.8) American Indian or 0 2 (0.6) 2 (0.3) Alaska Native Native Hawaiianor 0 0 0 Other Pacific Islander Other 1 (0.3) 4 (1.2) 5 (0.7) 0.3022Ethnicity, n (%) n 340 345 685 Hispanic or Latino 8 (2.4) 8 (2.3) 16(2.3) Not Hispanic or Latino 328 (96.5) 335 (97.1) 663 (96.8) NotReported/Unknown 4 (1.2) 2 (0.6) 6 (0.9) 0.9061 Age (years) n 340 345685 Mean (SD) 61.3 (15.34) 62.2 (15.12) 61.8 (15.23) Median 61.0 63.063.0 Min, Max 19, 97 19, 94 19, 97 0.3188 Categorical Age (years), n (%)n 340 345 685 18-45 55 (16.2) 52 (15.1) 107 (15.6) >45-65 145 (42.6) 141(40.9) 286 (41.8) >65 140 (41.2) 152 (44.1) 292 (42.6) 0.7574 >75 70(20.6) 75 (21.7) 145 (21.2) Height (cm) n 340 345 685 Mean (SD) 168.8(9.84) 168.5 (9.82) 168.7 (9.82) Median 169.0 168.0 169.0 Min, Max 137,196 135, 198 135, 198 0.7520 Weight (kg) n 340 345 685 Mean (SD) 77.70(17.203) 78.20 (18.029) 77.95 (17.613) Median 75.80 78.00 77.00 Min, Max 38.7, 136.0  28.0, 145.2  28.0, 145.2 0.8760 BMI (kg/m²) n 340 345 685Mean (SD) 27.25 (5.623) 27.48 (5.821) 27.37 (5.721) Median 26.23 26.5726.37 Min, Max 16.5, 50.6 12.6, 54.7 12.6, 54.7 0.5139 Renal Function(Local Lab), n (%) n 340 345 685 Normal renal function 164 (48.2) 180(52.2) 344 (50.2) [CrCl >80 mL/min] Mild renal impairment 119 (35.0) 112(32.5) 231 (33.7) [CrCl >50-80 mL/min] Moderate renal impairment 57(16.8) 53 (15.4) 110 (16.1) [CrCl 30-50 mL/min] Severe renal impairment0 0 0 0.5897 [CrCl <30 mL/min] PORT Score (actual) n 340 345 685 Mean(SD) 83.5 (15.98) 83.6 (15.19) 83.6 (15.57) Median 80.0 82.0 81.0 Min,Max  51, 130  53, 123  51, 130 0.5004 PORT Risk Class (actual) n 340 345685 I (0 ≤ Port Score ≤ 50) 0 0 0 II (51 ≤ Port Score ≤ 70) 45 (13.2) 49(14.2) 94 (13.7) III (71 ≤ Port Score ≤ 90) 204 (60.0) 202 (58.6) 406(59.3) IV (91 ≤ PortScore ≤ 130) 91 (26.8) 94 (27.2) 185 (27.0) V (PortScore ≥ 131) 0 0 0 0.9048

In the table above, Age is calculated from the date of birth to theinformed consent date. p-values for differences between treatment groupsare from Fisher's exact test (for categorical variables) or WilcoxonRank Sum test (for continuous variables). For each categoricalparameter, the denominator for the percentage is the number of subjectswho had that parameter assessed. PORT Score (actual) and PORT Risk Class(actual) are based on PORT score (derived/corrected) from CRF.

c) Efficacy Analysis

For all efficacy analyses, subject data was analyzed in the group towhich the subject was randomized.

The following table summarizes the primary analysis results for earlyclinical response determined 72-120 hours after the first infusion oftest article in the ITT population.

Compound 1 Moxifloxacin (N = 386) (N = 388) Difference Efficacy Outcomen (%) n (%) (95% CI) Clinical Success 313 (81.1) 321 (82.7) −1.6 (−7.1,3.8) Clinical Failure or 73 (18.9) 67 (17.3) Indeterminate ClinicalFailure 49 (12.7) 47 (12.1) Indeterminate 24 (6.2) 20 (5.2) CI =Confidence Interval. Difference was observed difference in EarlyClinical Success rate between the Compound 1 and moxifloxacin groups.95% CI was constructed based on the Miettinen and Nurminen methodwithout stratification. Percentages were based on the number of subjectsin each treatment group.

The early clinical success rates (at 72-120 hr) in the ITT population,for both Compound 1 and Moxifloxacin, are depicted in FIG. 2. See theleft most pair of bars. The data shows that the observed −1.6%difference in clinical success rate is well within the 10% margin ofstatistical non-inferiority between −7.1% and 3.8%, at 95% CI(Confidence Interval), and thus the primary efficacy point (for FDAapproval) is met.

For Early Clinical Response Efficacy Variable (Clinical Success,Clinical Failure or Indeterminate), an Indeterminate Response wasincluded in the denominator for the calculation of the percentage ofsubjects with a Clinical Success in the ITT population and thus, wasessentially considered as a Clinical Failure for the purpose of theprimary analysis.

The following table summarizes the overall clinical response at PTEvisit based on investigator assessment of the ITT and CE-PTEpopulations.

Popula- Efficacy Compound 1 Moxifloxacin Difference tion Outcome n (%) n(%) (95% CI) ^([1]) ITT (N = 386) (N = 388) Clinical 338 (87.6) 330(85.1) 2.5 (−2.4, Success 7.4) Clinical Failure 48 (12.4) 58 (14.9) orIndeterminate Clinical Failure 32 (8.3) 42 (10.8) Indeterminate 16 (4.1)16 (4.1) CE-PTE (N = 340) (N = 345) Clinical 316 (92.9) 312 (90.4) 2.5(−1.7, Success 6.8) Clinical Failure 24 (7.1) 33 (9.6) ^([1]) 95% CI wasconstructed based on the Miettinen and Nurminen method withoutstratification. CI = Confidence Interval; Difference was observeddifference in Overall Clinical Success rate at PTE between the Compound1 and Moxifloxacin groups. Overall Clinical Response at PTE was based onthe Investigator Assessment at the EOT and PTE visits. Percentages werebased on the number of subjects in each treatment group. EOT = End ofTreatment; PTE = Post Treatment Evaluation.

The overall clinical response rates at PTE visit based on investigatorassessment of the ITT population and the CE-PTE population, for bothCompound 1 and Moxifloxacin, are also depicted in FIG. 2. See the middle(ITT) and the right most (CE-PTE) pairs of bars. The data shows that theobserved 2.5% difference in overall clinical response rate in the ITTpopulation is within the 10% margin of statistical non-inferioritybetween −2.4% and 7.4%, at 95% CI (Confidence Interval); and that theobserved 2.5% difference in overall clinical response rate in the CE-PTEpopulation is within the 10% margin of statistical non-inferioritybetween −1.7% and 6.8%, at 95% CI (Confidence Interval). Thus thesecondary efficacy point (for FDA approval) is also met.

For Investigator's Assessment of Clinical Response at PTE EfficacyVariable (Clinical Success, Clinical Failure or Indeterminate in the ITTpopulation and Clinical Success and Clinical Failure in the CEpopulation), an Indeterminate Response was included in the denominatorfor the calculation of the percentage of subjects with a ClinicalSuccess in the ITT population and thus, was essentially considered aClinical Failure for the purpose of the primary analysis for the EMA.

To demonstrate the efficacy of Compound 1 was non-inferior tomoxifloxacin in the treatment of adults with CABP, the followinghypothesis were evaluated by analysis of the Clinical Success rates.

The null hypothesis and alternate hypothesis for the Early ClinicalResponse endpoint was assessed in the ITT population as follows:

H_(o): θ_(T)−θ_(C)≤−ΔH_(ai): θ_(T)−θ_(C)>−Δ

Where the clinical success rate for the Compound 1 regimen was θ_(T) andfor moxifloxacin is θ_(C). A was the non-inferiority (NI) margin and was0.10 (or 10%).

Similar null and alternative hypotheses can be set up with Δ of 0.10 forthe PTE endpoint. For the Early Clinical Response (FDA) endpoint, a2-sided 95% confidence interval (CI) approach for the difference ofclinical success rates (using the point estimate of the difference:Compound 1 response proportion minus moxifloxacin response proportion)was used to test for the NI of the Compound 1 arm compared to themoxifloxacin arm in the ITT population. The 95% CI was calculated usingthe unstratified method proposed by Miettinen and Nurminen (Statisticsin Medicine 4:213-226, 1985). Compound 1 is considered non-inferior tomoxifloxacin if the lower bound of the CI is greater than −0.10 (or−10%). This concept is expressed herein as “within 10% margin ofnon-inferiority.”

For Investigator's Assessment of Clinical Response at PTE (EMA) primaryefficacy analyses in both the ITT and CE populations, a 2-sided 97.5% CIapproach for the difference of clinical success rates (using the pointestimate of the difference: Compound 1 response proportion minusmoxifloxacin response proportion) was used to test for the NI of theCompound 1 arm compared to the moxifloxacin arm in those subjects with aPORT Risk Class of III or higher. The 97.5% CI was calculated using thestratified (for the randomization stratification factors) methodproposed by Miettinen and Nurminen. Compound 1 is considerednon-inferior to moxifloxacin if the lower bound of the CI is greaterthan −0.10 (or −10%).

Early Clinical Response and Investigator's Assessment of ClinicalResponse at PTE was tested separately and were not co-primary endpoints.The probability for approving an ineffective drug based on PTE efficacywas 1.25%, regardless of the result for the Early Clinical Responseendpoint and vice versa. An adjustment would only be required if winningon at least 1 endpoint would result in global approval which was not thecase here. In addition, no alpha adjustment was needed for theco-primary efficacy endpoints for the EMA (ITT and CE populations) sinceNI must be shown in both populations to conclude NI. Hence there was noadjustment for multiple endpoints.

Additional and sensitivity analyses of the primary efficacy outcomes(Early Clinical Response and Investigator's Assessment of ClinicalResponse at PTE) were performed. If the null hypothesis of inferiorityis rejected for the Early Clinical Response in the ITT population andthe observed success response proportion for Compound 1 is larger thanthe observed proportion for moxifloxacin, a formal statistical analysisof superiority would be conducted. If the lower limit of the 2-sided CIfor the treatment difference is greater than 0%, Compound 1 will beconsidered superior to moxifloxacin.

The primary efficacy outcome was also assessed separately across thestratification factors of PORT Risk Class, receipt of allowedantibacterial therapy in the 72 hours prior to study treatment andgeographic region stratum by treatment group. For each PORT Risk Classstratum, each prior antibacterial therapy stratum and each geographicregion stratum, a 2-sided 95% CI for the observed difference in EarlyClinical Response rates were calculated for the ITT population.Additional subgroup analyses of the primary efficacy outcome might havebeen conducted as descriptive analyses.

The following two tables summarize the overall clinical response at PTEvisit based on investigator assessment by PORT Risk Class in the ITTpopulation and the CE-PTE population, respectively.

Overall Clinical Response at PTE Visit based on Investigator Assessmentby PORT Risk Class in the ITT Population PORT Risk Class as RandomizedActual PORT Risk Class^([1]) Compound 1 Moxifloxacin Compound 1Moxifloxacin PORT Risk Class (N = 386) (N = 388) Difference (N = 386) (N= 388) Difference Efficacy Outcome n (%) n (%) (CI) n (%) n (%) (CI)PORT Risk Class II^([2]) 56 56 57 56 Clinical Success 45 (80.4) 47(83.9) −3.6 (−18.2, 11.0)^([4]) 47 (82.5) 47 (83.9) −1.5 (−15.7,12.8)^([4]) Clinical Failure or Indeterminate 11 (19.6) 9 (16.1) 10(17.5) 9 (16.1) Clinical Failure 6 (10.7) 7 (12.5) 5 (8.8) 7 (12.5)Indeterminate 5 (8.9) 2 (3.6) 5 (8.8) 2 (3.6) PORT Risk Class III/IV 330332 329 331 Clinical Success 293 (88.8) 283 (85.2) 3.5 (−1.6, 8.7)^([4])291 (88.4) 282 (85.2) 3.3 (−1.9, 8.5)^([4]) (−2.7, 9.3)^([5]) ClinicalFailure or Indeterminate 37 (11.2) 49 (14.8) 38 (11.6) 49 (14.8)Clinical Failure 26 (7.9) 35 (10.5) 27 (8.2) 35 (10.6) Indeterminate 11(3.3) 14 (4.2) 11 (3.3) 14 (4.2) PORT Risk Class III 241 232 227 216Clinical Success 217 (90.0) 201 (86.6) 3.4 (−2.4, 9.4)^([4]) 206 (90.7)190 (88.0) 2.8 (−3.0, 8.7)^([4]) Clinical Failure or Indeterminate 24(10.0) 31 (13.4) 21 (9.3) 26 (12.0) Clinical Failure 17 (7.1) 21 (9.1)16 (7.0) 18 (8.3) Indeterminate 7 (2.9) 10 (4.3) 5 (2.2) 8 (3.7) PORTRisk Class IV^([3]) 89 100 102 116 Clinical Success 76 (85.4) 82 (82.0)3.4 (−7.5, 14.0)^([4]) 85 (83.3) 93 (80.2) 3.2 (−7.4, 13.4)^([4])Clinical Failure or Indeterminate 13 (14.6) 18 (18.0) 17 (16.7) 23(19.8) Clinical Failure 9 (10.1) 14 (14.0) 11 (10.8) 17 (14.7)Indeterminate 4 (4.5) 4 (4.0) 6 (5.9) 6 (5.2) The overall clinicalresponse rates at PTE visit based on investigator assessment of the ITTpopulation limited to actual PORT Risk Class III/IV subjects, for bothCompound 1 and Moxifloxacin, are depicted in FIG. 3. See the left pairsof bars. The data shows that the observed 3.3% difference in overallclinical response rate is within the 10% margin of statisticalnon-inferiority between −2.7% and 9.3%, at 97.5% CI (ConfidenceInterval). Thus one of the co-primary efficacy point (for EMA approval)is met.

Overall Clinical Response at PTE Visit based on Investigator Assessmentby PORT Risk Class in the CE-PTE Population PORT Risk Class asRandomized Actual PORT Risk Class^([1]) Compound 1 Moxifloxacin Compound1 Moxifloxacin PORT Risk Class (N = 340) (N = 345) Difference (N = 340)(N = 345) Difference Efficacy Outcome n (%) n (%) (CI) n (%) n (%) (CI)PORT Risk Class II 44 48 45 49 Clinical Success 41 (93.2) 44 (91.7) 1.5(−11.2, 13.9)^([4]) 43 (95.6) 44 (89.8) 5.8 (−6.1, 18.1)^([4]) ClinicalFailure 3 (6.8) 4 (8.3) 2 (4.4) 5 (10.2) PORT Risk Class III/IV 296 297295 296 Clinical Success 275 (92.9) 268 (90.2) 2.7 (−1.9, 7.3)^([4]) 273(92.5) 268 (90.5) 2.0 (−2.6, 6.6)^([4]) (−3.2, 7.4)^([5]) ClinicalFailure 21 (7.1) 29 (9.8) 22 (7.5) 28 (9.5) PORT Risk Class III 216 212204 202 Clinical Success 202 (93.5) 193 (91.0) 2.5 (−2.7, 7.8)^([4]) 191(93.6) 186 (92.1) 1.5 (−3.6, 6.9)^([4]) Clinical Failure 14 (6.5) 19(9.0) 13 (6.4) 16 (7.9) PORT Risk Class IV 80 85 91 94 Clinical Success73 (91.3) 75 (88.2) 3.0 (−6.8, 12.8)^([4]) 82 (90.1) 82 (87.2) 2.9(−6.7, 12.5)^([4]) Clinical Failure 7 (8.8) 10 (11.8) 9 (9.9) 12 (12.8)^([1])Actual PORT Risk Class was based on PORT score (derived/corrected)from CRF. ^([2])Also includes 4 subjects with an actual PORT Risk Classof I. ^([3])Also includes 1 subject with an actual PORT Risk Class of V.^([4])Unstratified 95% CI was constructed based on the Miettinen andNurminen method without stratification. ^([5])Stratified 97.5% CI wasadjusted for prior antibiotics use (yes and no) constructed based on theMiettinen and Nurminen method with stratification, usingCochran-Mantel-Haenszel weights as stratum weights. CI = ConfidenceInterval; PTE = Post Treatment Evaluation. Difference was observeddifference in Overall Clinical Success rate at PTE between the Compound1 and Moxifloxacin groups. Percentages were based on the number ofsubjects in each treatment group within each PORT Risk Class. Theoverall clinical response rates at PTE visit based on investigatorassessment of the CE-PTE population, limited to patients with PORTIII/IV CABP, for both Compound 1 and Moxifloxacin, are also depicted inFIG. 3. See the right pairs of bars. The data shows that the observed2.0% difference in overall clinical response rate is within the 10%margin of non-inferiority between -3.2% and 7.4%, at 97.5% CI(Confidence Interval). Thus another of the co-primary efficacy point(for EMA approval) is also met.

Sensitivity analyses include: conducting an adjusted analysis of theprimary efficacy outcome based on the randomized stratum and separately,based on the stratum the subject actually belongs, and conducting ananalysis where all subjects with an Indeterminate response areconsidered Clinical Successes.

d) Analysis of Secondary Variables

The number and percentage of subjects classified as a Clinical Success,Clinical Failure and Indeterminate by the Investigator's Assessment atPTE in the ITT and CE populations (by definition subjects with anIndeterminate response were excluded from the CE population) werecalculated for each treatment group. A 2-sided unadjusted 95% CI wasconstructed for the observed difference in the clinical success rateusing the method of Miettinen and Nurminen. For Investigator'sAssessment of Clinical Response at PTE in the ITT and CE populations the95% CI was for descriptive purposes only and no conclusion of NI wasmade. The number and percentage of subjects in each treatment group ineach response category for Early Clinical Response was presented for themicrolTT population. The number and percentage of subjects who wereclassified as a Clinical Success and Clinical Failure by theinvestigator at the PTE visit in ME population were calculated.Two-sided unadjusted 95% CI was constructed for the observed differencein the clinical success rates using the method of Miettinen andNurminen.

The number and percentage of subjects with an Early Clinical Response ofsuccess and an Investigator's Assessment of Clinical Response at PTE ofClinical Success by pathogen was provided in the microlTT and MEpopulations. All-cause mortality (ACM) at 15 and 30 days after the firstdose of test article was summarized in the ITT population. Subjects whowere lost to follow-up were considered deceased for this analysis. A2-sided unadjusted 95% CI for the observed difference in mortality rateswas calculated for ACM.

e) Analysis of Additional Efficacy Variables

Additional efficacy analyses were conducted to support the efficacyfindings of the primary and secondary outcomes. CIs were determined fordescriptive purposes, but no conclusions of NI were made. The number andpercentage of subjects classified as a Clinical Success, ClinicalFailure and Indeterminate by the Investigator's Assessment at EOT in theITT and CE populations (by definition subjects with an Indeterminateresponse were excluded from the CE population) were calculated. A2-sided unadjusted 95% CI was constructed for the observed difference inthe Clinical Success rate using the method of Miettinen and Nurminen.The number and percentage of subjects with stabilization of vital signsand clinical signs/laboratory findings associated with CABP at 72-120hours post first dose of test article were presented by treatment groupin the ITT population. These include body temperature (no fever orhypothermia), SBP (>90 mm Hg), heart rate (<90 bpm), RR (<20breaths/minute), PaO₂ (≥60 mm Hg by ABG or oxygen saturation ≥90% bypulse oximetry), physical exam findings (no findings of pulmonaryconsolidation), WBC count (<12,000 cells/mm³ or ≥4,000 cells/mm³) orimmature neutrophils (<15%). A summary (number and percentage ofsubjects) of the assessment of clinical signs and symptoms of CABP ateach time point throughout the study were presented by treatment groupin the ITT population. The number and percentage of subjects withresolution of signs and symptoms present at Screening (back to pre-CABPstatus) were also provided by study visit. The number and percentage ofsubjects with no worsening of clinical symptoms of CABP and with theabsence of new symptoms of CABP were provided by treatment group in theITT population.

The per-subject and per-pathogen microbiologic outcomes were providedfor the microlTT and ME populations at the EOT and PTE visits. Forexample, the table below summarizes the overall clinical success at PTEvisit in the microlTT population, based on investigator assessment bybaseline pathogen from blood specimens, respiratory specimens, urinaryantigen tests, and/or serology.

Compound 1 Moxifloxacin (N = 204) (N = 182) Clinical Success ClinicalSuccess Baseline Pathogen N1 n (%) N1 n (%) Gram-Positive Bacteria(aerobes) 61 52 (85.2) 56 49 (87.5) Streptococcus pneumoniae ^([1]) 4337 (86.0) 34 31 (91.2) MDRSP 7 7 (100.0) 6 6 (100.0) PSSP 26 23 (88.5)22 21 (95.5) Macrolide Resistant 10 10 (100.0) 5 5 (100.0)Staphylococcus aureus 11 8 (72.7) 11 9 (81.8) MRSA 0 0 1 1 (100.0) MSSA11 8 (72.7) 10 8 (80.0) Beta Hemolytic Streptococcus 2 2 (100.0) 3 1(33.3) Streptococcus agalactiae 2 2 (100.0) 3 1 (33.3) Streptococcusanginosus 0 0 1 0 Streptococcus constellatus 1 1 (100.0) 1 1 (100.0)Streptococcus gordonii 0 0 1 1 (100.0) Streptococcus mitis 3 3 (100.0) 53 (60.0) Streptococcus mitis group 1 1 (100.0) 2 2 (100.0) Streptococcusoralis 1 1 (100.0) 0 0 Streptococcus parasanguinis 0 0 2 1 (50.0)Streptococcus salivarius 1 1 (100.0) 3 3 (100.0) Streptococcus sanguinis0 0 2 1 (50.0) Streptococcus sanguis ii 2 2 (100.0) 0 0 Gram-NegativeBacteria 79 67 (84.8) 68 55 (80.9) (aerobes) Acinetobacter baumanii 0 01 1 (100.0) Acinetobacter baumanii complex 0 0 1 0 Acinetobacter junii 11 (100.0) 0 0 Acinetobacter lwoffii 1 1 (100.0) 0 0 Citrobacter braakii0 0 1 1 (100.0) Citrobacter freundii 0 0 1 1 (100.0) Enterobactercloacae 2 2 (100.0) 4 3 (75.0) Escherichia coli 6 4 (66.7) 7 4 (57.1)Haemophilus haemolyticus 1 1 (100.0) 0 0 Haemophilus influenzae 32 26(81.3) 16 16 (100.0) Haemophilus parahaemolyticus 2 2 (100.0) 2 1 (50.0)Haemophilus parainfluenzae 18 15 (83.3) 17 13 (76.5) Klebsiella oxytoca1 0 4 4 (100.0) Klebsiella pneumoniae 13 10 (76.9) 13 11 (84.6)Moraxella catarrhalis 4 4 (100.0) 1 1 (100.0) Morganella morganii 1 1(100.0) 0 0 Neisseria meningitidis 1 1 (100.0) 1 1 (100.0) Proteusmirabilis 2 1 (50.0) 2 2 (100.0) Pseudomonas aeruginosa 3 2 (66.7) 5 5(100.0) Pseudomonas putida 0 0 1 1 (100.0) Serratia marcescens 1 1(100.0) 0 0 Stenotrophomonas maltophilia 0 0 2 1 (50.0) AtypicalPathogens 118 109 (92.4) 106 97 (91.5) Mycoplasma pneumoniae 70 66(94.3) 57 50 (87.7) Chlamydophila pneumoniae 28 25 (89.3) 28 25 (89.3)Legionella pneumophila ^([2]) 37 35 (94.6) 37 36 (97.3) Gram-negativeorganisms 0 0 1 0 (anaerobes) Prevotella oris 0 0 1 0 ^([1])Overalltabulation of Streptococcus pneumoniae includes identification fromurinary antigen only which will not have susceptibility data. ^([2])Legionella pneumophila may be detected from culture, serology and/orurinary antigen test. Subjects with the same pathogen isolated frommultiple specimens are counted only once for that pathogen. Subjectswith the same pathogen from a blood specimen, respiratory specimen,urinary antigen test, and/or serology are counted only once for thatpathogen. Subjects are counted only once in the overall tabulations ifthey have more than one respective pathogen at baseline. PTE = PostTreatment Evaluation. N = Number of subjects in the microITT population.N1 = Number of subjects with the specific baseline pathogen, n = Numberof subjects in the specific category. Percentages are based on thenumber of subjects with the specific baseline pathogen.

For selected pathogens that have 10 or more isolates in the Compound 1treatment arm, data was extracted and shown in FIG. 4.

Two-sided unadjusted 95% CIs were provided for the difference inper-subject microbiological favorable outcome rates. A concordanceanalysis of Early Clinical Response and Investigator's Assessment ofClinical Response at PTE in the ITT analysis set were also presented.

f) Safety Outcome Measures

Safety variables included the incidence rate of AEs, change in vitalsigns, ECG parameters and laboratory test results obtained during thecourse of the study.

Summary tables were provided for all treatment-emergent adverse events(TEAEs), defined as an AE with a start date and time on or after thefirst dose of test article. AEs were summarized by presenting the numberand percentage of subjects having each TEAE for each treatment group bysystem organ class (SOC) and preferred term (PT). Additional tabulationsprovided summaries by SOC and PT of subjects experiencing SAEs, severeTEAEs, TEAEs judged to be related to test article, TEAEs leading todiscontinuation of test article, TEAEs leading to dose interruption oftest article, and TEAEs of special interest.

The following table provides an overview of Adverse Events (AEs) withinthe safety population.

Compound 1 Moxifloxacin All Subjects (N = 382) (N = 388) (N = 770)Parameter n (%) n (%) n (%) Total Number of AEs 434 520 954 Total Numberof TEAEs 376 461 837 Subjects with at Least One, n (%) Adverse Events(AE) 170 (44.5) 200 (51.5) 370 (48.1) TEAE 157 (41.1) 188 (48.5) 345(44.8) Drug-Related TEAE 39 (10.2) 69 (17.8) 108 (14.0) Severe TEAE 25(6.5) 26 (6.7) 51 (6.6) Serious TEAE 23 (6.0) 26 (6.7) 49 (6.4)Drug-Related Serious TEAE 2 (0.5) 2 (0.5) 4 (0.5) Serious TEAE Leadingto Death* 8 (2.1) 4 (1.0) 12 (1.6) TEAE Leading to PrematureDiscontinuation of Test Article 21 (5.5) 27 (7.0) 48 (6.2) TEAE Leadingto Premature Discontinuation of Study 7 (1.8) 9 (2.3) 16 (2.1) TEAELeading to Dose Interruption 0 0 0 Serious TEAEs Leading to PrematureDiscontinuation of Test Article 10 (2.6) 11 (2.8) 21 (2.7) Subjects whodied, n (%) 8 (2.1) 4 (1.0) 12 (1.6) *All deaths occurred inpatients >65 years of age; frequency consistent with recently completedCABP studies. Percentages are based on the Safety population. A TEAE isdefined as an AE occurring after first dose of active test article. AE =Adverse Events; TEAE = Treatment-Emergent Adverse Event.

The following table provides a summary of selected TEAEs by PreferredTerm within the safety population. With few exceptions, the selectedTEAEs have a rate of occurrence of at least about 2% in the Compound 1treatment group.

Omadacycline Moxifloxacin All Subjects (N = 382) (N = 388) (N = 770)Preferred Term (PT) n (%) n (%) n (%) Subjects with at Least One TEAE157 (41.1) 188 (48.5) 345 (44.8) Alanine Aminotransferase Increased 14(3.7) 18 (4.6) 32 (4.2) Hypertension 13 (3.4) 11 (2.8) 24 (3.1)Gamma-Glutamyltransferase Increased 10 (2.6) 8 (2.1) 18 (2.3) Insomnia10 (2.6) 8 (2.1) 18 (2.3) Vomiting 10 (2.6) 6 (1.5) 16 (2.1)Constipation 9 (2.4) 6 (1.5) 15 (1.9) Nausea 9 (2.4) 21 (5.4) 30 (3.9)Aspartate Aminotransferase Increased 8 (2.1) 14 (3.6) 22 (2.9) Headache8 (2.1) 5 (1.3) 13 (1.7) . . . Diarrhoea 4 (1.0) 31 (8.0) 35 (4.5)Dyspepsia 4 (1.0) 2 (0.5) 6 (0.8) Clostridium Difficile Colitis 0 1(0.3) 1 (0.1) Clostridium Difficile Infection 0 6 (1.5) 6 (0.8)Pseudomembranous Colitis 0 1 (0.3) 1 (0.1) Coding of Preferred Term isbased on MedDRA Version 17.1. Percentages are based on the Safetypopulation. A TEAE is defined as an AE occurring after the first dose ofactive test article. If a subject has more than one TEAE that codes tothe same MedDRA category, the subject is counted only once. PTs aresorted by decreasing frequency within the Compound 1 column.

Thus, it is apparent that gastrointestinal (GI) adverse events (AEs,including vomiting, nausea, diarrhea, and dyspepsia) associated withtreatment with Compound 1 are mild.

The following table summarizes selected TEAEs that led todiscontinuation of treatment.

Compound 1 Moxifloxacin All Subjects System Organ Class (SOC) (N = 382)(N = 388) (N = 770) Preferred Term (PT) n (%) n (%) n (%) Subjects withat Least One 21 (5.5)  27 (7.0)  48 (6.2)  TEAE Leading to Study DrugDiscontinuation Gastrointestinal Disorders 2 (0.5) 2 (0.5) 4 (0.5)Vomiting 2 (0.5) 0 2 (0.3) Nausea 1 (0.3) 0 1 (0.1) Diarrhoea 0 1 (0.3)1 (0.1) Dyspepsia 0 1 (0.3) 1 (0.1) Coding of System Organ Class (SOC)and Preferred Term (PT) was based on MedDRA Version 17.1. Percentageswere based on the Safety population. A TEAE was defined as an AEoccurring after the first dose of active test article. If a subject hadmore than one TEAE that coded to the same MedDRA category, the subjectwas counted only once. SOC terms were sorted alphabetically, then PTswere sorted within each SOC term by decreasing frequency within theCompound 1 column.

The following variables were analyzed descriptively as vital signs:

-   -   Vital signs (systolic and diastolic BP, pulse rate, body        temperature, RR) including change from Screening by visit    -   Clinically notable vital signs (meeting predefined criteria as        specified in the SAP) by visit

Electrocardiogram data (RR interval, PR interval, QRS interval,Corrected QT interval [QTc], QTc Bazzett's Correction Formula [QTcB],and QTc Fridericia's Correction Formula [QTcF]) was summarizeddescriptively at each scheduled evaluation and for the overall worstpost-Screening value. Changes from Screening at each visit were alsoprovided. An outlier analysis was conducted based on the worstpost-Screening value.

The following variables were analyzed descriptively for laboratorytests:

-   -   Laboratory variables by visit    -   Change from Screening of laboratory variable by visit    -   Clinically notable laboratory values (meeting predefined        criteria specified in SAP) by visit

g) PK

Population PK analysis was conducted to characterize PK parameters. Apopulation PK data set including subjects with 1 or more quantifiedCompound 1 concentration determinations was constructed from the datesand times of the doses and blood samples along with all thebioanalytical determinations and subject background information. If theactual date or time for a blood sample or dose was missing, the relatedbioanalytical determination of the PK concentration was excluded fromall analyses. Compound 1 concentrations below the limit ofquantification were treated as missing data in summary statistics andfor the calculation of PK parameters.

Variables including age (years), body weight (kg), gender, andrace/ethnicity along with other covariates previously determined to beimportant were incorporated into the population PK database. Based onthe subjects in the population analysis data set, descriptive summariesat Screening for these variables were reported. Outliers may be excludedfrom the analysis. These were determined by a scatter plot of theobserved concentration versus time post dose and reported. Thedistribution of the number of samples contributed per subject to themodel-based analysis was tabulated. Also, simple summary descriptivestatistics for the concentration of samples by study day or week wascomputed.

Regarding population PK modeling, results from Phase 1 studies indicatedthat Compound 1 PK was linear and that following iv infusion, plasmaconcentration-time profiles show a 3-compartmental disposition.Therefore, the probable structural PK model would be a 3-compartmentmodel with zero order input for iv infusion and first order input for poadministration. This PK model contained the parameters clearance, volumeof distribution, bioavailability and absorption rate constant. Theassociated population models were nonlinear mixed-effects models. Thepopulation model added random effects and covariates for the PKparameters in order to recognize differences among individuals andsimilarities across observations corresponding to the same subject. Atthe time of the population modeling, previously reported structural PKmodels were considered first. A residual error model combining additiveerror and proportional error was also considered. Simplifications (e.g.,fewer random effects, or an alternative residual error model) may beappropriate if the diagnostics for the model suggest false convergence.Additional covariates were investigated graphically (gender,race/ethnicity, age) as part of the model diagnostics and some may beretained in the final model and additional ones in a competing model todeliver estimates of arguably insignificant effects. Scatter plots ofthe observed concentrations versus population-estimated and individuallyestimated concentrations were used as part of the overall assessment ofthe overall quality of the fit. During modeling, the broad principlesoutlined by the FDA were followed. The individual model-based exposuremeasures at steady state (area under the Concentration/Time curve[AUC_(0-24,ss)], time to maximum plasma concentration [T_(max,ss)],maximum plasma concentration [_(Cmax,ss)]) were computed and summarized.

The relationship between Compound 1 exposure and response (efficacy andsafety) was examined as appropriate for the data. A population PK modelwas used to calculate individual subject AUCs and, subsequently,possible AUC/MIC breakpoints.

Example 4 a Phase I, Randomized, Double-Blind, 3-Period, Crossover Studyto Evaluate Safety, Tolerability, and Pharmacokinetics of Multiple OralDoses of Omadacycline or Placebo in Healthy Adult Subjects

The primary objective of this study was to assess and compare thepharmacokinetics (PK) of 300-, 450-, and 600-mg doses of oralomadacycline administered daily over 5 days. The secondary objective ofthe study was to evaluate the safety and tolerability of multiple dosesof omadacycline in healthy adult subjects.

For the treatment of CABP, the then anticipated therapeutic daily oraldose (excluding any loading dose) was 300 mg. For potential futurestudies, or for administration of a loading dose using the oralformulation, it is possible that a daily dose higher than 300 mg couldbe used to achieve omadacycline concentrations sufficient to treattarget bacteria in the organs/tissues of interest. One early clinicalstudy evaluated single oral doses of omadacycline up to 600 mg, but nostudies have evaluated multiple daily doses higher than 300 mg. Thisstudy was designed to obtain data on the safety, tolerability, andpharmacokinetics (dose linearity and proportionality) of multiple oraldoses of omadacycline at daily doses higher than 300 mg. Placebo groupswere included as a reference to minimize potential bias in assessingtolerability.

Multiple daily oral doses of 300, 450, and 600 mg omadacycline orplacebo were chosen to be administered in this study. The lowest dose of300 mg had been evaluated in multiple dose studies and had been welltolerated; this daily dose has also been studied in Phase 3 studies inABSSSI. Single oral doses up to 600 mg were administered in capsules tohealthy adult subjects in 1 early clinical study and were determined tohave an acceptable safety profile. There was some increased incidence ofGI AEs at oral doses of 400 mg or greater, though events were typicallymild (none were severe), and it is possible that some of these eventsmay have been related to the oral formulation. Multiple daily doses ofup to 600 mg using the final optimized tablet formulation ofomadacycline were expected to have acceptable safety profiles, but thiswas important to assess in a small carefully controlled Phase 1 studybefore evaluating these doses in larger clinical studies.

Thus the study was designed as a Phase 1, randomized, double-blind,3-period, crossover study in healthy adult subjects. The study consistedof a screening period (Day −21 through Day −2), 3 baseline periods (Day−1 of each period), 3 treatment periods (Day 1 through Day 6 of eachperiod), and a study completion visit (within 6 to 10 days after thelast dose of study drug in Period 3). There was a washout of at least 5days between the last dose in one period and the first dose in the nextperiod. Subjects were confined to the study site from Day −1 of Period 1until discharge on Day 6 of Period 3, after the 24-hour blood sampling,urine sampling, and safety assessments were completed. Subjects returnedto the study center 6 to 10 days after the last dose of study drug inPeriod 3 for the study completion visit.

Subject Selection

Healthy, non-smoking, male and female subjects were eligible forparticipation in the study if they were between 18 and 55 years of age(inclusive), weighed ≥50 kg, had a body mass index between 18 and 30kg/m2 (inclusive), met all eligibility criteria during screening(performed within 21 days before dosing in Period 1) and at baseline(Day −1) for Period 1, and provided written informed consent. Healthstatus was determined by past medical history, clinical laboratorytests, vital signs (oral body temperature, systolic blood pressure,diastolic blood pressure, and heart rate), 12-lead electrocardiogram(ECG), and physical examination at screening. Eligibility criteriaincluded ability to swallow up to 4 tablets in succession.

Subjects were excluded from participation in the study for priortreatment with omadacycline, recent use of other investigational drugs;ECG abnormalities; inability to tolerate oral medications; pregnancy orbreastfeeding; use of tobacco products, prescription drugs, herbalsupplements, or over-the-counter medications or intake of xanthine(e.g., caffeine)—containing food or beverages within a specified timeframe before study initiation; blood loss/donation; low hemoglobinlevels; high creatinine or blood urea nitrogen levels; urinaryobstruction/difficulty voiding; positive alcohol or drug test;hypersensitivity or allergy to any tetracycline; signs of liver diseaseor liver injury; significant illness within 2 weeks of study initiation;any planned medical intervention that might interfere with the study; ora history of diseases or medical conditions as specified in the studyprotocol.

Study Design

On Day 1 through Day 5 of each period, subjects received once-daily,after a fast of 6 hours, one of the following treatments (omadacyclineor placebo) according to the randomization schedule:

-   -   A. 300 mg omadacycline (2×150-mg tablets)    -   AP. Placebo for 300 mg omadacycline (2×placebo tablets)    -   B. 450 mg omadacycline (3×150-mg tablets)    -   BP. Placebo for 450 mg omadacycline (3×placebo tablets)    -   C. 600 mg omadacycline (4×150-mg tablets)    -   CP. Placebo for 600 mg omadacycline (4×placebo tablets)

All doses of study drug were administered in the morning with no food ordrink except for water at least 6 hours prior to dosing. Subjects thenhad no food or drink except water for at least 2 hours after dosing andno dairy products, antacids, or multivitamins for 4 hours after dosing.

Before the dosing, subjects underwent screening evaluations to determineeligibility within 21 days before dosing in Period 1. Subjects were thenadmitted to the clinical site on the day before dosing (Day −1 ofPeriod 1) for baseline evaluations. Before dosing on Day 1 of Period 1,up to 30 subjects (24 omadacycline, 6 placebo) were randomly assigned to1 of 3 treatment sequences using a Latin Square design as presented inthe following table:

Number of Period Period Period Sequence Subsequence Subjects 1 2 3 1 1A8 omadacycline A C S 1B 2 placebo AP CP BP 2 2A 8 omadacycline B A C 2B2 placebo BP AP CP 3 3A 8 omadacycline C B A 3B 2 placebo CP BP AP

About ten subjects were randomly assigned to each sequence. Placebo wasadministered to 2 subjects in each sequence as a reference to assesstolerability. Subjects assigned to omadacycline received omadacyclineduring all 3 periods and at all tested dose levels. Subjects assigned toplacebo received placebo during all 3 periods. Investigators andsubjects were blinded to whether the subject was receiving omadacyclineor placebo.

Study Asessment

1. Plasma Pharmacokinetics

Serial blood samples for pharmacokinetic (PK) analysis of omadacyclinewere collected at specified time points through 24 hours after dosing onDay 1 and Day 5 of each period. Specifically, blood samples for PKassessments of omadacycline were collected from all subjects at thefollowing time points: before dosing (predose) and at 0.5, 1, 1.5, 2,2.5, 3, 4, 6, 8, 12, 16, and 24 hours after dosing on Day 1 and Day 5 ineach period. The 24-hour blood sample for Day 1 was collected prior todosing on Day 2 for each period.

Non-compartmental PK parameters were determined on Days 1 and 5 of eachperiod from plasma omadacycline concentration and actual time data usingPhoenix® WinNonlin® (Certara, Princeton, N.J.) Version 6.2.1., includingarea under the plasma concentration versus time curve (AUC) from time 0to 24 hours after dosing (AUC₀₋₂₄), AUC from time 0 to the lastquantifiable concentration (AUC_(iast)), maximum observed plasmaconcentration (C_(max)), time to reach maximum observed plasmaconcentration (T_(max)), terminal elimination half-life (T_(1/2)),terminal phase rate constant (λ_(z)) and the accumulation factor (Rac)of AUC₀₋₂₄ and C_(max).

Subjects that received omadacycline and had at least one evaluable PKparameter were included in the PK analysis population; however, subjectsmay have been excluded from the PK population if they missed doses, haddiarrhea, or had vomiting at or before a time equal to twice the medianT_(max).

2. Urine Pharmacokinetics

Urine samples were collected from a subset of subjects at specifiedintervals on Day 5 of Period 2 and on Day 1 and Day 5 of Period:predose, 0 to 4, 4 to 8, 8 to 12, and 12 to 24 hours after dosing. The12 to 24-hour interval urine sample for Day 1 was collected prior todosing on Day 2. Urine samples were only collected from a subset ofsubjects because analysis of urine PK was added by an amendment to thestudy protocol after the study was already underway.

The following urine PK parameters were determined from urineomadacycline concentration and collection interval data using SASVersion 9.2: renal clearance (CLr), fraction of the dose excretedunchanged in urine from 0 to 24 hours after dosing (Fe₀₋₂₄), and amountof drug excreted unchanged in urine over 24 hours after dosing(Ae_(t1-t2)). Additional parameters Ae₀₋₄, Ae₄₋₈, Ae₈₋₁₂, Ae₁₂₋₂₄, andAe₀₋₂₄ were also calculated.

3. Safety and Tolerability

Safety assessments include monitoring of adverse events (AEs), clinicallaboratory test results, vital sign measurements, 12-leadelectrocardiogram (ECG) results, and physical examination findings. Allrandomly assigned subjects who received at least one dose of any studydrug (omadacycline or placebo) were included in the safety analysispopulation. Adverse events were coded by preferred term and system organclass using MedDRA Version 17.1.

Safety and tolerability were assessed by the monitoring and recording ofAEs, clinical laboratory test results (hematology, serum chemistry, andurinalysis), vital sign measurements (oral body temperature, systolicblood pressure, diastolic blood pressure, and HR), 12-lead ECG results,and physical examination findings.

Statistical Analysis for Pharmacokinetic Study:

Individual plasma and urine concentration and time deviation data werepresented in data listings. Plasma and urine concentration data weresummarized by day and time point or interval for each treatment usingdescriptive statistics (number of subjects, mean, SD, coefficient ofvariation [CV], median, minimum, and maximum). Concentrations that werebelow the limit of quantification (BLQ) were treated as zero in theplasma and urine concentration descriptive statistics summaries. Meanand individual plasma concentration versus time profiles were presentedin figures on both linear and semilogarithmic scales.

Noncompartmental PK parameters were determined from plasma concentrationand actual time data using Phoenix® WinNonlin® (Certara, Princeton,N.J.) Version 6.2.1 or higher. Urine PK parameters were determined fromurine concentration and collection interval data using SAS Version 9.2or higher. All further statistical analyses were performed using SAS®software (SAS Institute, Cary, N.C.), Version 9.2.

For the PK analysis, BLQ values were treated as zero with the exceptionthat a BLQ value between 2 quantifiable concentrations were set asmissing. Missing concentrations were treated as missing from the PKparameter calculations. If consecutive BLQ concentrations were followedby quantifiable concentrations in the terminal phase, thoseconcentrations after BLQ concentrations were treated as missing.

The individual PK parameters were presented in data listings.Descriptive statistics (number of subjects, mean, SD, CV, median,minimum, and maximum) were calculated for the PK parameter estimatesafter dosing on Day 1 and Day 5 of each period (e.g., AUC₀₋₂₄,AUC_(last), C_(max), T_(max), T_(1/2), and Rac [Day 5 only] from plasmaconcentrations; CLr, Fe₀₋₂₄, and Ae₀₋₂₄ from urine concentrations).Geometric means were included for AUC₀₋₂₄, AUC_(last), and C_(max).

A linear mixed-effect model (SAS PROC MIXED) with treatment (A, B, andC), sequence (1A, 2A, and 3A), and treatment period as fixed effects andsubject nested within sequence as a random effect were fitted to thenatural log-transformed dose normalized PK parameters AUC₀₋₂₄/Dose,AUC_(last)/Dose, and C_(max)/Dose after dosing on Day 1 and Day 5 ofeach period for use in estimation of effects and construction ofconfidence intervals (CIs). Point estimates and 90% CIs for differenceson the log scale were exponentiated to obtain estimates for the ratiosof geometric means and respective 90% CIs on the original scale. Noadjustment was made for multiplicity.

Dose linearity across all 3 dose levels was assessed by fittingomadacycline C_(max), AUC_(last), and AUC₀₋₂₄ after both the Day 1 andDay 5 doses to a power model (10): ln(PK)=a +b×ln(Dose)+error, where PKwas the PK parameter, a was the intercept and b was the slope. Theestimates of slope b were reported along with the corresponding 2-sided90% CIs.

For statistical analysis of accumulation of omadacycline, a linearmixed-effect model with day as a fixed effect and subject as randomeffect was fitted to the natural log-transformed C_(max) and AUC₀₋₂₄ toconstruct 90% CIs for Day 5 compared with Day 1 (at each dose levelseparately).

Results

a. Demographics, Baseline Characteristics, and Disposition of StudySubjects

Of the 33 subjects enrolled in the study, 26 were assigned to receiveomadacycline and 7 were assigned to receive placebo. Demographic andbaseline characteristics were generally similar between omadacycline andplacebo treatment groups (Table 4-1) and across all omadacyclinetreatment sequences (data not shown). The majority of subjects in thestudy were white (57.6%) and male (81.8%). The overall mean age ofsubjects was 36.9 years, with a range of 21 to 55 years.

TABLE 4-1 Demographics and Baseline Characteristics of Subjects in theStudy^(a) Omadacycline Placebo Overall (N = 26) (N = 7) (N = 33) Age,years Mean (±SD) 35.6 (±10.4) 41.9 (±11.6) 36.9 (±10.8) Min, max 21, 5525, 53 21, 55 Sex, n (%) Male 21 (80.8) 6 (85.7) 27 (81.8) Female 5(19.2) 1 (14.3) 6 (18.2) Race, n (%) White 15 (57.7) 4 (57.1) 19 (57.6)Black or African American 9 (34.6) 3 (42.9) 12 (36.4) Asian 2 (7.7) 0 2(6.1) Ethnicity, n (%) Hispanic or Latino 10 (38.5) 3 (42.9) 13 (39.4)Not Hispanic or Latino 16 (61.5) 4 (57.1) 20 (60.6) Height, cm Mean(±SD) 173.12 (±9.17) 172.89 (±4.31) 173.07 (±8.32) Min, max 155.2, 192.4165.6, 177.4 155.2, 192.4 Weight, kg Mean (±SD) 78.67 (±10.33) 83.77(±4.80) 79.75 (±9.60) Min, max  62.7, 101.4 76.7, 90.4  62.7, 101.4 Bodymass index, kg/m² Mean (±SD) 26.25 (±2.72) 28.04 (±1.45) 26.63 (±2.59)Min, max 19.4, 29.8 25.8, 29.9 19.4, 29.9 ^(a)Results for SafetyPopulation

All 33 subjects received at least one dose of study drug (omadacyclineor placebo) and were included in the safety analysis population.Twenty-five of the 26 omadacycline-treated subjects (96.2%) wereincluded in the PK analysis population (one subject was excluded fromthis population due to vomiting after dosing). Four omadacycline-treatedsubjects (15.4%) and one placebo-treated subject (14.3%) discontinuedthe study; these early discontinuations were due to treatment-emergentadverse events (TEAEs) in 4 subjects (see below); in addition oneomadacycline-treated subject was lost to follow-up. Thus, 22 subjectsreceived all 5 doses of 300-, 450-, and 600-mg omadacycline and 6subjects received all 5 doses of placebo in Periods 1, 2, and 3. Thesesubjects were considered to have completed the study.

b. Plasma Pharmacokinetics

At all tested omadacycline dose levels on both Day 1 and Day 5 of each5-day treatment period, mean plasma omadacycline concentrations peaked2.5 hours after dosing (T_(max)) and omadacycline was measurable inplasma for up to 24 hours after dosing (the last sampling time) (FIG. 5and Table 4-2).

TABLE 4-2 Plasma Pharmacokinetic Parameters of Omadacycline by Dose onDays 1 and 5 of Dosing^(a) Omadacycline Dose Day 1 Day 5 300 mg 450 mg600 mg 300 mg 450 mg 600 mg Parameter (n = 25) (n = 24) (n = 24) (n =23) (n = 24) (n = 23) Mean AUC₀₋₂₄, 6644.8 8976.5 10020.5 9267.2 13366.716420.3 ng · h/mL (25.3) (26.6) (25.7) (26.8) (26.0) (27.1) (CV) MeanC_(max), ng/mL 648.8 874.2 954.5 808.8 1077.3 1305.5 (CV) (24.0) (26.6)(23.2) (25.9) (25.0) (26.6) Mean T_(max), h 2.50 2.50 2.51 2.50 2.502.50 (Min, max) (1.50, 3.00) (1.50, 3.00) (1.00, 3.00) (1.00, 3.00)(1.50, 4.00) (2.00, 4.00) Mean T_(1/2), h 13.66 13.45 13.03 15.49 16.8316.75 (CV) (12.5)^(b) (12.9)^(c) (11.8)^(c) (10.7)^(d) (8.1)^(c)(6.8)^(d) ^(a)Results for Pharmacokinetic Population ^(b)n = 24 (T_(1/2)was not estimable for 1 subject) ^(c)n = 23 (T_(1/2) was not estimablefor 1 subject) ^(d)n = 21 (T_(1/2) was not estimable for 2 subjects) CV,coefficient of variation Note: One 300 mg omadacycline subject and one600 mg omadacycline subject were excluded from the Day 5 summary due tovomiting before reaching the pharmacokinetic steady state on Day 5Omadacycline total exposure (AUC₀₋₂₄ and AUC_(last)) and peakconcentrations (C_(max)) increased with increasing omadacycline dose(300 vs 450 vs 600 mg) on both Day 1 and Day 5, and were higher on Day 5than on Day 1 for corresponding doses (FIG. 5 and Table 4-2). The meanhalf-life of omadacycline in plasma (T_(1/2)) was similar across the 3tested dose levels, ranging from 13.03 to 13.66 hours on Day 1 and from15.49 to 16.83 hours on Day 5 (Table 4-2). Between-subject variabilityin systemic omadacycline exposure was low and was similar at all threetested dose levels, with coefficients of variation (CVs) ranging from23.2% to 26.6% for C_(max), AUC₀₋₂₄, and AUC_(last) on Day 1 and from25.0% to 27.1% for C_(max), AUC₀₋₂₄, and AUC_(last) on Day 5 (Table4-2).

Although omadacycline AUC₀₋₂₄, AUC_(last) and C_(max) increased withincreasing omadacycline dose, the observed increases in exposure wereless than dose proportional on both days of analysis (Tables 4-2 and4-3).

TABLE 4-3 Statistical Analysis of Dose-Normalized OmadacyclinePharmacokinetic Parameters on Days 1 and 5 of Dosing^(a) Ratio ofGeometric Treatment Geometric 90% CI of Parameter Treatment N LS MeansComparison LS Means (%) Ratio (%) Day 1 AUC₀₋₂₄/Dose 300 mg 25 21.32 (ng· h/mL/mg) 450 mg 24 18.64 450/300 87.44 (77.41, 98.77) 600 mg 24 16.18600/450 86.79 (76.71, 98.20) 600/300 75.89 (67.20, 85.71) C_(max)/Dose300 mg 25 2.09 (ng/mL/mg) 450 mg 24 1.81 450/300 86.71 (76.17, 98.71)600 mg 24 1.54 600/450 85.26 (74.76, 97.23) 600/300 73.92 (64.95, 84.14)Day 5 AUC₀₋₂₄/Dose 300 mg 23 30.09 (ng · h/mL/mg) 450 mg 24 28.83450/300 95.82  (90.39, 101.59) 600 mg 23 26.46 600/450 91.78 (86.58,97.30) 600/300 87.95 (82.96, 93.25) Cmax/Dose 300 mg 23 2.62 (ng/mL/mg)450 mg 24 2.32 450/300 88.58 (83.19, 94.32) 600 mg 23 2.11 600/450 90.72(85.20, 96.60) 600/300 80.36 (75.47, 85.58) ^(a)Results forPharmacokinetic Population ANOVA analysis; see Materials and Methods fordetails CI, confidence interval; LS, least squares Note: One 300 mgomadacycline subject and one 600 mg omadacycline subject were excludedfrom the Day 5 statistical analysis due to vomiting before reaching thepharmacokinetic steady state on Day 5

Statistical analyses showed that with an increase in dose from 300 mg to600 mg, omadacycline exposure (based on dose-normalized AUC₀₋₂₄) on Day1 was 76% of that predicted if exposure were perfectly dose-proportional(Table 4-3); on Day 5, the observed increase in omadacycline exposurewas 88% of predicted (Table 4-3). Analysis of C_(max) values similarlydemonstrated that omadacycline concentrations were dose-linear, but lessthan dose-proportional in this study (Tables 4-2 and 4-3).

Statistical analyses also revealed accumulation of omadacycline inplasma following once-daily dosing for 5 consecutive days. Depending ondose, accumulation ratios between Day 5 and Day 1 ranged from 1.40 to1.62 for AUC₀₋₂₄ and from 1.24 to 1.35 for C_(max) (data not shown).These findings are consistent with the long half-life of omadacycline inplasma.

The above data showed that mean concentrations of omadacycline peaked at2.5 hours and remained measurable up to 24 hours (the last testedtimepoint) at all omadacycline dosing levels (300, 450, and 600 mg). OnDay 5, mean steady state exposure (AUC₀₋₂₄) in subjects dosed with300-mg omadacycline was 9267 ng·h/mL, which is consistent with resultsof previous studies with 300 mg oral dosing. Both AUC₀₋₂₄ and C_(max)increased with increasing dose and were nearly, but somewhat less than,dose-proportional (74%-88% of expected). This was the case on both Day 1and Day 5 of dosing. Due to its relatively long half-life (mean=˜13 h onDay 1, ˜16 h on Day 5), omadacycline accumulated in plasma over thecourse of 5 consecutive days of dosing. Thus, at all tested dose levels,systemic exposure on Day 5 was ˜50% higher than on Day 1. This degree ofaccumulation is also consistent with that observed following multipleonce-daily dosing of IV or oral formulations of omadacycline in earlypharmacology studies.

In terms of systemic exposure, this study showed that omadacyclineplasma concentrations on Day 1 of 450-mg dosing were similar to those onDay 5 of 300-mg dosing (mean AUC₀₋₂₄=8976.5 and 9267.2 ng·h/mL,respectively). For indications in which the therapeutic dosing regimenincorporates 300 mg daily oral dosing, these data support a strategy ofusing an initial oral “loading dose” of 450 mg once-daily for 1-2 days,followed by 300-mg once-daily oral dosing. Such a strategy couldpotentially eliminate the need for an IV phase of treatment.

c. Urine Pharmacokinetics

Because urine sample collection and PK analysis were added to the studyby protocol amendment after the study was underway, only a limitednumber of samples were evaluated (samples from 9 subjects on Day 5 ofPeriod 2 and samples from 8 subjects on Day 1 and Day 5 of Period 3).While this sample size was too small to make meaningful comparisonsbetween omadacycline dose groups, the results of the analysis didprovide an overall indication of partial omadacycline renal clearanceand urinary excretion.

For all omadacycline dose groups, the mean fraction of the dose excretedunchanged in urine from 0 to 24 hours after dosing (Fe₀₋₂₄) ranged from−5% to ˜7% on Day 1 and from −7% to ˜9% on Day 5. Renal clearance (CLr)ranged from 2.8 to 4.2 L/h on Day 1 and from 2.4 to 3.3 L/h on Day 5(Table 4-4).

TABLE 4-4 Urine Pharmacokinetic Parameters of Omadacycline by Dose onDays 1 and 5 of Dosing^(a) Omadacycline Dose Day 1 Day 5 300 mg 450 mg600 mg 300 mg 450 mg 600 mg Parameter^(b) (n = 2) (n = 3) (n = 1) (n =3) (n = 5) (n = 4) Ae₀₋₂₄ (mg) 20.37 (8.3) 25.06 (16.8) 31.96 26.14(14.6)  30.81 (33.0)  51.82 (14.8)  Fe₀₋₂₄ (%) 6.79 (8.3)   5.57 (16.8)5.33 8.71 (14.6) 6.85 (33.0) 8.64 (14.8) CLr (L/h) 3.01 (11.4) 2.80(9.6) 4.17 3.28 (27.2) 2.38 (34.9) 3.05 (19.9) ^(a)Results forPharmacokinetic Population ^(b)Mean (CV) CV, coefficient of variationNote: One 600 mg omadacycline subject was excluded from the summary dueto vomiting before reaching the pharmacokinetic steady state on Day 5

Urine PK analyses in a subset of subjects provided preliminaryindications of partial renal clearance and urinary excretion ofomadacycline. On Day 5, depending on dose level, ˜7% to ˜9% of theadministered oral dose was excreted unchanged in the urine over 24hours. This represents approximately 20% to 25% of the absorbed dosesince it is known that the absolute bioavailability of the tabletformulation used in this study is 35%. Presence of unchangedomadacycline in the urine suggests that it may be useful in urinarytract infections, an indication that is currently being explored.

d. Safety and Tolerability

Overall, 12 of the 33 subjects in the safety population reported a totalof 36 TEAEs during the study (Table 4-5).

TABLE 4-5 Summary of Treatment-Emergent Adverse Events^(a) OmadacyclineDose Omadacycline Placebo 300 mg 450 mg 600 mg Overall Overall (n = 26)(n = 24) (n = 24) (n = 26) (n = 7) n (%) of subjects with: Any TEAE 5(19.2) 3 (12.5) 6 (25.0) 10 (38.5) 2 (28.6) Treatment-related TEAE 4(15.4) 2 (8.3) 6 (25.0) 9 (34.6) 1 (14.3) Most frequent TEAEs (seenin >1 study subject), n (%) Nausea 2 (7.7) 1 (4.2) 4 (16.7) 6 (23.1) 0Vomiting 2 (7.7) 0 1 (4.2) 3 (11.5) 0 Diarrhea 0 0 2 (8.3) 2 (7.7) 0Dizziness 2 (7.7) 0 1 (4.2) 3 (11.5) 0 ALT increased 0 1 (4.2) 1 (4.2) 2(7.7) 0 TEAEs leading to early discontinuation of study drug, n (%) All1 (3.8) 1 (4.2) 1 (4.2) 3 (11.5) 1 (14.3) Nausea 1 (3.8) 0 0 1 (3.8) 0Vomiting 1 (3.8) 0 0 1 (3.8) 0 ALT increased 0 1 (4.2) 0 1 (3.8) 0Lipase increased 0 0 1 (4.2) 1 (3.8) 0 Syncope 0 0 0 0 1 (14.3)^(b)^(a)Results for Safety Population ^(b)vasovagal syncope following ablood draw ALT, alanine aminotransferase; TEAE, treatment-emergentadverse event

TEAEs were reported by 38.5% of subjects that received omadacycline and28.6% of subjects that received placebo. The highest percentage of TEAEswas classified as gastrointestinal (GI) disorders. The most frequentlyreported TEAE was nausea, which occurred in <7.7% of the omadacycline300 and 450 mg dose groups and 16.7% of the 600 mg group. All of theTEAEs reported in this study were either mild or moderate in severity.There were no serious TEAEs (SAEs) reported during the study. Foursubjects experienced TEAEs leading to study discontinuation, includingone subject at each of the 3 omadacycline dose levels and 1 subject inthe placebo group.

There were no clinically significant findings in analyses of vital signmeasurements, physical examination, ECG results, hematology orurinalysis parameters. Serum chemistry analyses showed that betweenbaseline and Day 5 of each dosing period, the median change in alanineaminotransferase (ALT) concentration was −2.0, 5.0 and 19.5 IU/L insubjects dosed with 300, 450 and 600 mg omadacycline, respectively. Thecorresponding changes in placebo groups ranged from −5.0 to −1.0 IU/L.No substantial changes in median aspartate aminotransferase (AST),bilirubin or other serum chemistry parameters were noted. The highestindividual ALT value was 150 IU/L (2.7-fold above the upper limit ofnormal [ULN]), which occurred in a subject who first received 450 mgomadacycline in Period 1 then 300 mg in Period 2 and then wasdiscontinued due to the liver enzyme changes; this subject's bilirubinvalues remained within the normal range at all time points assessed.

The plasma PK findings indicate that higher systemic drug exposure canbe achieved by increasing the amount of omadacycline administered perdose during once-daily oral dosing, but that the exposure benefit is notdose-proportional. Moreover, increasing omadacycline dosing beyond acertain point appears to have adverse effects in terms of safety andtolerability. While multiple doses of 300, 450, and 600 mg were allgenerally well-tolerated in this study (all TEAEs were either mild ormoderate in severity), there were some differences between the doses.The frequency of treatment-related TEAEs did not increase with anincrease in omadacycline dose from 300 to 450 mg (15.4% vs 8.3%), butsuch events were more frequent with 600 mg (25.0%). Within the mostfrequent class of TEAEs, GI disorders, nausea occurred with incidence atleast 9% higher for the 600 mg dose level than for the lower doses, andthe only 2 reports of diarrhea occurred with 600 mg. In addition, serumchemistry analyses showed a small but notable dose-dependent increase inmedian ALT concentrations. While no individual ALT values exceeded3-fold above the ULN, the higher median ALT at 600 mg suggests anincreased chance of more significantly elevated serum transaminaselevels with this dose. Based on these findings, for situations in whichan oral dose above 300 mg may be beneficial, 450 mg was identified asthe oral dose most likely to provide higher omadacycline exposure withfavorable safety and tolerability.

In summary, this phase 1 study investigated the pharmacokinetics (PK)and safety/tolerability of multiple oral omadacycline doses higher than300 mg. Using a 3-period crossover design, healthy adults wererandomized to receive omadacycline (300-, 450- and 600-mg in variablesequence; n=26) or placebo (n=7) once daily for 5 consecutive days perperiod. In plasma, omadacycline maximum concentration and total exposureincreased with increasing dose, but were less than dose-proportional(74% to 88% of expected). The kinetics of omadacycline plasmaaccumulation were similar between dose levels; exposure on Day 5 was˜50% higher than on Day 1. Omadacycline plasma concentrations on Day 1of 450 mg dosing were similar to those on Day 5 of 300 mg dosing. UrinePK analyses indicated partial renal clearance and urinary excretion ofunchanged omadacycline. All doses were generally well-tolerated. Theseresults support the use of once-daily 450-mg oral omadacycline as partof the oral only dosing regimen, such as using once-daily 450-mg oralomadacycline (either one or two doses) as loading dose before steppingdown to once-daily 300-mg oral omadacycline, or in a dosing regimenusing once-daily 450-mg oral omadacycline throughout the treatment.

What is claimed is:
 1. A method of treating Community-Acquired BacterialPneumonia (CABP) in a subject in need of treatment thereof, comprisingadministering to said subject 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline or a salt thereof according to the followingdosing regimen: (1) three intravenous doses of about 100 mg each,administered 12 hours apart, followed by, (2) one or more intravenousdoses of about 100 mg each, each administered 24 hours following theimmediate preceding intravenous dose, followed by, (3) optionally, oneoral dose of about 300 mg, administered in the morning and 12-24 hrsafter the immediate preceding intravenous dose, followed by, (4)optionally, one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding oral dose, suchthat said subject is treated.
 2. A method of treating Community-AcquiredBacterial Pneumonia (CABP) in a subject in need of treatment thereof,comprising administering to said subject 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline or a salt thereof according to the followingdosing regimen: (1) three intravenous doses of about 100 mg each,administered 12 hours apart, followed by, (2) optionally, one or moreintravenous doses of about 100 mg each, each administered 24 hoursfollowing the immediate preceding intravenous dose, followed by, (3)optionally, one oral dose of about 300 mg, administered in the morningand 12-24 hrs after the immediate preceding intravenous dose, followedby, (4) optionally, one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding oral dose, suchthat said subject is treated.
 3. A method of treating Community-AcquiredBacterial Pneumonia (CABP) in a subject in need of treatment thereof,comprising administering to said subject 9-[(2,2-dimethyl-propylamino)-methyl]-minocycline or a salt thereof according to the followingdosing regimen: (1) three intravenous doses of about 100 mg each,administered 12 hours apart, followed by, (2) one or more intravenousdoses of about 100 mg each, each administered 24 hours following theimmediate preceding intravenous dose, followed by, (3) one or more oraldoses of about 300 mg each, each administered 24 hours following theimmediate preceding dose, such that said subject is treated.
 4. A methodof treating Community-Acquired Bacterial Pneumonia (CABP) in a subjectin need of treatment thereof, comprising administering to said subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three intravenous dosesof about 100 mg each, administered 12 hours apart, followed by, (2)optionally, one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, followed by, (3) one or more oral doses of about 300 mg each, eachadministered 24 hours following the immediate preceding dose, such thatsaid subject is treated.
 5. The method of claim 3 or 4, wherein step (2)consists of one intravenous dose of about 100 mg of said subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or salt thereof.
 6. Amethod of treating Community-Acquired Bacterial Pneumonia (CABP) in asubject in need of treatment thereof, comprising administering to saidsubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) three intravenousdoses of about 100 mg each, administered 12 hours apart, followed by,(2) one or more intravenous doses of about 100 mg each, eachadministered 24 hours following the immediate preceding intravenousdose, such that said subject is treated.
 7. The method of any one ofclaims 1-6, wherein the steps are completed within 7 days, 8 days, 9days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17days, 18 days, 19 days, or 20 days.
 8. The method of any one of claims1-6, wherein the steps are completed within 7-14 days.
 9. The method ofclaim 8, wherein the steps are completed within 7-10 days.
 10. Themethod of claim 8, wherein the steps are completed within 11-14 days.11. The method of claim 8, wherein the steps are completed within 10days.
 12. The method of any one of claims 1-11, wherein the number ofdays of IV dosing is 3-10 days.
 13. The method of claim 12, wherein thenumber of days of IV dosing is 3-6 days.
 14. The method of claim 12,wherein the number of days of IV dosing is 7-10 days.
 15. The method ofclaim 12, wherein the number of days of IV dosing is 5 days.
 16. Themethod of any one of claims 1-15, comprising one or more oral doses, andwherein the number of days of IV dosing is 4-7 days.
 17. The method ofclaim 16, wherein the number of days of IV dosing is 4-5 days.
 18. Themethod of claim 16, wherein the number of days of IV dosing is 6-7 days.19. The method of claim 16, wherein the number of days of IV dosing is 5days.
 20. The method of any one of claims 16-19, wherein the number ofdays of oral dosing is 1-7 days.
 21. The method of claim 20, wherein thenumber of days of oral dosing is 1-4 days.
 22. The method of claim 20,wherein the number of days of oral dosing is 5-7 days.
 23. The method ofclaim 20, wherein the number of days of oral dosing is 5 days.
 24. Themethod of claim 16, wherein the number of days of IV dosing is 5 days,and the number of days of oral dosing is 5 days.
 25. A method oftreating Community-Acquired Bacterial Pneumonia (CABP) in a subject inneed of treatment thereof, comprising administering to said subject9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a salt thereofaccording to the following dosing regimen: (1) three oral doses of about300-450 mg each, administered 12 hours apart, followed by, (2)optionally, one or more oral doses of about 300-600 mg each, eachadministered 24 hours following the immediate preceding oral dose, suchthat said subject is treated.
 26. The method of claim 25, wherein eachoral dose is about 300 mg.
 27. The method of claim 25, wherein each oraldose is about 450 mg.
 28. The method of claim 25, wherein each oral dosein step (1) is about 300 mg.
 29. The method of claim 25, wherein eachoral dose in step (1) is about 450 mg.
 30. The method of claim 25, 28,or 29, wherein each oral dose in step (2) is about 300 mg.
 31. Themethod of claim 25, 28, or 29, wherein each oral dose in step (2) isabout 450 mg.
 32. The method of claim 25, 28, or 29, wherein each oraldose in step (2) is about 600 mg.
 33. The method of claim 25, whereinthe first two oral doses of step (1) are each 300 mg, and the last oraldose of step (1) is about 300, 450, or 600 mg.
 34. The method of claim25, wherein the first two oral doses of step (1) are each 450 mg, andthe last oral dose of step (1) is about 300, 450, or 600 mg.
 35. Amethod of treating Community-Acquired Bacterial Pneumonia (CABP) in asubject in need of treatment thereof, comprising administering to saidsubject 9-[(2,2-dimethyl-propyl amino)-methyl]-minocycline or a saltthereof according to the following dosing regimen: (1) one or twoonce-daily oral dose(s) of about 450-600 mg (administered 24 hrs apartfor two once-daily oral doses), followed by, (2) one or more oral dosesof about 300-600 mg each, each administered 24 hours following theimmediate preceding oral dose, such that said subject is treated. 36.The method of claim 35, wherein said dosing regimen is: (1) one or twoonce-daily oral dose(s) of about 450 or 600 mg (administered 24 hrsapart for two once-daily oral doses), followed by, (2) one or more oraldoses of about 300 mg each, each administered 24 hours following theimmediate preceding oral dose.
 37. The method of claim 35, wherein saiddosing regimen is: (1) two once-daily oral doses of about 450 mg,administered 24 hrs apart, followed by, (2) one or more oral doses ofabout 300 mg each, each administered 24 hours following the immediatepreceding oral dose.
 38. The method of any one of claims 25-37, whereinthe steps are completed within 7 days, 8 days, 9 days, 10 days, 11 days,12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days,20 days, or 21 days.
 39. The method of any one of claims 25-37, whereinthe steps are completed within 7-14 days, within 7-10 days, within 11-14days, or within 10 days.
 40. The method of any one of claims 1-39,wherein said CABP is caused by Staphylococcus aureus includingmethicillin-resistant Staphylococcus aureus (MRSA), Streptococcuspneumoniae including penicillin-resistant Streptococcus pneumoniae(PRSP), Haemophilus influenzae, Moraxella catarrhalis, Klebsiellapneumoniae, Legionella pneumophila, Chlamydophila pneumoniae, Mycoplasmapneumoniae, Chlamydophila psittaci, Coxiella burnetii, Escherichia coli,or a combination thereof.
 41. The method of claim 40, wherein saidStreptococcus pneumoniae is penicillin-resistant Streptococcuspneumoniae (PRSP), macrolide-resistant Streptococcus pneumoniae,cephalosporin-resistant Streptococcus pneumoniae, or multidrug-resistantStreptococcus pneumoniae (MDRSP).
 42. The method of any one of claims1-39, wherein said CABP is caused by intracellular pathogens, such asLegionella pneumophila, Mycoplasma pneumoniae, Chlamydophila pneumoniae,Chlamydophila psittaci, Coxiella burnetii, or a combination thereof. 43.The method of any one of claims 1-39, wherein said CABP is caused byHaemophilus parainfluenzae.
 44. The method of any one of claims 1-43,wherein said subject is a human.
 45. The method of any one of claims1-44, wherein each of said oral dose is administered independently astwo 150-mg tablets.
 46. The method of any one of claims 1-45, whereineach of said intravenous dose is administered continuously over about 30minutes (e.g., at least 30 minutes and not more than 45 minutes). 47.The method of any one of claims 1-46, wherein said dosing regimen has aclinical success rate that is within 10% (or 12.5%) margin ofnon-inferiority compared to moxifloxacin administered as 400 mgintravenous dose once every 24 hours for three or more days, followed byone or more doses of 400 mg oral doses of moxifloxacin once every 24hours.
 48. The method of any one of claims 1-47, wherein said subjectexperience improvement, at day 3 to day 5 after step (1), in at leasttwo symptoms selected from: chest pain, frequency or severity of cough,amount of productive sputum, and difficulty breathing, wherein saidsymptoms are evaluated on a four-point scale of absent, mild, moderate,and severe, and wherein improvement is at least a one-point improvementfrom baseline to the assessment at said day 3 to day 5 (e.g., fromsevere to moderate, from moderate to absent, or from mild to absent).49. The method of any one of claims 1-47, wherein said subject, at day 3to day 5 after step (1), experience improvement in at least two symptomsand no worsening in any of the symptoms selected from: chest pain,frequency or severity of cough, amount of productive sputum, anddifficulty breathing, and improvement in at least one vital signselected from: body temperature, blood pressure, heart rate, andrespiratory rate.
 50. The method of any one of claims 1-49, wherein thesubject undergoes fasting overnight, with no food or drink except waterfor at least 6 hours, just before step (3) dosing (if present), andwherein the subject continues fasting after step (3) dosing, with nofood for 2 hours, and no dairy products for 4 hours.
 51. The method ofany one of claims 1-50, wherein said salt is a tosylate salt.
 52. Themethod of any one of claims 1-51, which method has a clinical successrate of about 70%-100%.
 53. The method of claim 52, wherein saidclinical success rate is about 75-95%, about 80-95%, about 75-90%, about80-90%, about 75-85%, about 80-85%, about 85-90%, about 90-95%, about80-82%, or about 81%.
 54. The method of claim 53, wherein said clinicalsuccess rate is about 75-85%, observed at about 72-120 hours after theadministration of the first intravenous dose.
 55. The method of claim54, wherein said clinical success rate is about 80-82%, or 80% or 81%.56. The method of claim 53, wherein said clinical success rate isobserved at about 5-10 days after the last dose of treatment (e.g.,equivalent to a time for post treatment evaluation in clinicallyevaluable population, or CE-PTE; or in ITT population).
 57. The methodof claim 56, wherein said clinical success rate is about 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, or97%.
 58. The method of any one of claims 1-57, wherein said subject hasCABP categorized as PORT Risk Class II.
 59. The method of claim 58,wherein said method has a clinical success rate of about 70-100%, about75-96%, about 75-90%, about 80-83%, about 82%, about 80-96%, about90-96%, or about 95%.
 60. The method of claim 59, wherein said clinicalsuccess rate is about 75-85%, or about 90-100%, observed at about 5-10days after the last dose of treatment.
 61. The method of claim 60,wherein said clinical success rate is about 82%, or about 95%.
 62. Themethod of any one of claims 1-57, wherein said subject has CABPcategorized as PORT Risk Class III.
 63. The method of claim 62, whereinsaid method has a clinical success rate of about 80-100%, about 85-95%,about 90-95%, about 90-91%, or about 93-94%.
 64. The method of claim 63,wherein said clinical success rate is about 85-100%, observed at about5-10 days after the last dose of treatment.
 65. The method of claim 64,wherein said clinical success rate is about 90-91%, or about 93-94%. 66.The method of any one of claims 1-57, wherein said subject has CABPcategorized as PORT Risk Class IV.
 67. The method of claim 66, whereinsaid method has a clinical success rate of about 70-100%, about 75-95%,about 80-95%, about 83-85%, or about 90-91%.
 68. The method of claim 67,wherein said clinical success rate is about 80-95%, observed at about5-10 days after the last dose of treatment.
 69. The method of claim 68,wherein said clinical success rate is about 83-85%, or about 90-91%. 70.The method of any one of claims 1-57, wherein said subject has CABPcategorized as PORT Risk Class III or IV.
 71. The method of claim 70,wherein said method has a clinical success rate of about 75-100%, about85-95%, about 85-90%, about 88-89%, about 90-95%, or about 92-93%. 72.The method of claim 71, wherein said clinical success rate is about85-95%, observed at about 5-10 days after the last dose of treatment.73. The method of claim 72, wherein said clinical success rate is about88-89%, or about 92-93%.
 74. The method of any one of claims 1-73,wherein gastrointestinal (GI) adverse events (AEs) associated withtreatment of said subject are mild.
 75. The method of any one of claims1-73, wherein GI adverse events (AEs) associated with treatment of saidsubject do not result in discontinuation of therapy with said method.76. The method of any one of claims 1-75, wherein treatment of saidsubject (1) does not result in increased risk of C. difficile (e.g., C.difficile colitis and Pseudomembranous colitis) infection in saidsubject, or (2) does not substantially disrupting gut microbiome in saidsubject.
 77. The method of claim 76, wherein said subject is at risk of,or is predisposed to, developing a C. difficile infection.
 78. Themethod of claim 77, wherein said subject has recently been treated withone or more antibiotics (such as broad spectrum antibiotics), has hadsurgery of the gastrointestinal tract, has a disease of the colon (suchas an inflammatory bowel disease or colorectal cancer), has a kidneydisease, has a weakened immune system; is on chemotherapy, haspreviously had C. difficile infection, is 65 years or older, takesproton-pump inhibitors, or is living in an environment that predisposessaid subject to developing C. difficile infection (such as in ahospital, a nursing home, or an assisted living facility).